Substituted pyrrolidines as factor xia inhibitors for the treatment thromboembolic diseases

ABSTRACT

The present invention provides compounds of the general formula (I), their salts and N-oxides, and solvates and prodrugs thereof (wherein the substituents are as defined in the description). The compounds of the general formula (I) are inhibitors of factor XIa, and are useful in the prevention of and/or therapy for thromboembolic diseases.

TECHNICAL FIELD

The present invention relates to a series of pyrrolidine derivativeswhich are useful as inhibitors of factor XIa.

Thus, the present invention relates to a compound of formula (I):

(wherein all symbols have the same meanings as described hereinafter) ora pharmaceutically acceptable salt thereof, an N-oxide thereof, asolvate thereof, or a prodrug thereof, use of such compounds intreatment and/or prevention of a thromboembolic disease and processesfor the preparation of said compounds.

BACKGROUND OF THE INVENTION

Thromboembolism is an important cause of morbidity and mortality. Itoccurs when a blood clot breaks free and is carried by the blood streamto obstruct a blood vessel at another site. Thromboembolic diseaseincludes venous thromboembolism, for example deep vein thrombosis orpulmonary embolism, arterial thrombosis, stroke and myocardialinfarction.

Thromboembolic diseases may be treated using anticoagulants. Oneapproach has been to target the inhibition of factor XIa (FXIa). FactorXIa is a plasma serine protease involved in the regulation of bloodcoagulation. Factor XIa is an activated form of factor XI, which isactivated by factor XIIa, thrombin, and it is also autocatalytic. FXIais a component of the “contact pathway” and activates factor IX byselectively cleaving arg-ala and arg-val peptide bonds. Factor IXa, inturn, activates factor X. The safety of this target is supported by theobservations that FXI deficiency in humans (hemophilia C) results in amild bleeding disorder. In addition to this, the efficacy and sideeffects of this target have been shown using experimental thrombosis andbleeding models in mice lacking FXI, and in baboons and rabbits treatedwith anti-FXI neutralizing antibodies. These results suggest that FXIainhibitors will show a potent anti-thrombotic effect without bleeding.Therefore, factor XIa is an attractive target for anti-thrombotictherapy without the side effect of bleeding.

It has been described in Patent literature 1 that compound of formula(A):

wherein A^(A) represents a 5- to 12-membered heterocycle, etc.; L_(1A)represents —CH═CH—, etc.; R^(11A) represents benzyl, etc.; M^(A)represents imidazolyl, etc; are useful as selective inhibitors of factorXIa or dual inhibitors of FXIa and plasma kallikrein.

Furthermore, it has been described in Patent literature 2 that acompound of formula (B-I):

wherein A^(B) represents a 5- to 12-membered heterocycle, etc.; L_(1B)represents —CH═CH—, etc.; R^(11B) represents benzyl, etc.; R^(3B)represents phenyl, etc.; R^(4B) represents chlorine, etc.; R^(8aB)represents hydrogen, etc; or formula (B-II):

wherein M^(B) represents pyridyl, etc.; and the other symbols have thesame meanings as described above; inhibit factor XIa and/or plasmakallikrein.

Furthermore, it has been described in Patent literature 3 that acompound of formula (C):

wherein D^(C) represents C10 cycloalkyl or 10-membered heterocycloalkyl,etc.; -L^(C)-E^(C)-G^(C)-J^(C)-represents —C—C—C—C, etc.; R^(3C)represents hydrogen, etc.; R^(4C) represents mono- or bicyclicheteroaryl, etc.; R^(5C) represents hydrogen, etc.; R^(13C) representshydrogen, etc.; M^(C) represents phenyl, etc.; are useful as inhibitorsof factor Xa.

Furthermore, it has been described in Patent literature 4 that acompound of formula (D):

wherein ring B^(D) represents phenyl, etc.; W^(D) represents —NH₂, etc.;Z^(1D) represents 5 to 7-membered monocyclic, etc.; L^(D) represents—NH—CO—, etc.; R^(1D) and R^(2D) independently represents (i) hydrogenor (ii) are taken together to form a five-to-seven membered fullysaturated heterocycle, etc.; R^(5D) and R^(6D) independently representshydrogen, etc.; R^(7D) represents —COOH, etc.; R^(8D) representshydrogen, etc.; (R^(27D))_(mD) represents —COOH, etc.; are useful asinhibitors of factor VIIa, factor IXa, factor FXIa, tryptase, andurokinase.

Furthermore, it has been described in Patent literature 5 that acompound of formula (E):

wherein A^(E) represents aryl substituted by carboxyl, etc.; R^(3E)represents hydrogen, etc.; X^(E) represents oxygen, etc.; V^(E)represents nitrogen, etc.; W^(E) represents carbon, etc.; Z^(E)represents —CO—, etc.; R^(E) represents aryl substituted by —C(═NH)NH₂,etc.; T^(E) represents C₂₋₆ alkylene, etc.; (Y^(1E))_(pE) representsheterocyclo substituted by —SO₂-Me, etc.; are useful as anti-viralagent, however, it is not reported that the compound represented byformula (E) has factor XIa inhibitory activity.

Furthermore, it has been described in Patent literature 6 that acompound of formula (F):

wherein ring X^(F) represents N-containing ring, etc.; A₁ ^(F)represents a bond, etc.; A₂ ^(F) represents Aryl, etc.; R₁ ^(F), R₂^(F), R₃ ^(F), R₄ ^(F) and R₅ ^(F) independently represents hydrogen,etc.; are useful as inhibitors of Apoptosis Proteins.

[Patent literature 1] WO2007070826

[Patent literature 2] WO2008076805

[Patent literature 3] WO2007131982

[Patent literature 4] WO2002037937

[Patent literature 5] WO2008064218

[Patent literature 6] WO2009152824

DISCLOSURE OF THE INVENTION

It is desirable to find new compounds which may be more effective intreating thromboembolic diseases. Advantageous compounds desirably havegood inhibitory and selectivity for factor XIa.

The present inventors have made extensive studies to find a compoundthat can become a therapeutic agent for thromboembolic diseases. As aresult, we have found that the object is achieved by a compoundrepresented by formula (I), a salt thereof an N-oxide thereof, a solvatethereof, or a prodrug thereof (hereinafter, which may be abbreviated tocompounds of the present invention) have good inhibitory and selectivityfor factor XIa and then we have completed the present invention.

Namely, the present invention relates to the following aspects:

(1) A compound represented by formula (I):

-   -   wherein Cyc A represents C3-C8 cycloalkyl, 5- to 10-membered        heterocycloalkyl, C6-C10 aryl or 5- to 10-membered heteroaryl;    -   Cyc B represents C3-C8 cycloalkyl, 5- to 10-membered        heterocycloalkyl, C6-C10 aryl or 5- to 10-membered heteroaryl;    -   Cyc C represents C3-C8 cycloalkyl, 5- to 10-membered        heterocycloalkyl, C6-C10 aryl or 5- to 10-membered heteroaryl;    -   each R¹ may be the same or different and represents (1) C6-C10        aryl, (2) 5- to 10-membered heteroaryl, (3) C6-C10 aryl or 5- to        10-membered heteroaryl substituted with 1 to 5 groups selected        from halogen, C1-4 alkyl, C1-4 alkoxy, —C1-4 alkylene-C1-4        alkoxy, CN, —COOH, —COO—C1-4 alkyl, —CO—NH₂, —OCONH₂,        —OCONH—C1-4 alkyl, —CONH—C1-4 alkyl, —NHCOO—C1-4 alkyl and        —NHCO—C1-4 alkyl, (4) —C(═NH)NH₂, (5) —NH—C(═NH)NH₂, (6) C1-4        alkyl, (7) C2-4 alkenyl, (8) C2-4 alkynyl, (9) —C1-4        alkylene-NH₂, (10) C1-4 alkoxy, (11) CN, (12) —CO—C1-4        alkyl, (13) halogen or (14) —R¹⁰—C(═NR¹¹)NR¹²R¹³;    -   wherein R¹⁰ represents (1) a bond or (2) NH;    -   R¹¹, R¹² and R¹³ each independently represents (1) hydrogen, (2)        OH, (3) C1-4 alkyl, (4) C2-4 alkenyl, (5) C2-4 alkynyl, (6) C1-4        alkoxy, (7) —C1-4 alkylene-C1-4 alkoxy, (8) —CO—C1-4 alkyl, (9)        —COO—C1-4 alkyl, (10) —OCO—C1-4 alkyl, (11) —CO—R¹⁴, (12)        —COO—R¹⁵ or (13) —OCO—R¹⁶, with the proviso that R¹¹, R¹² and        R¹³ do not all simultaneously represent hydrogen;    -   wherein R¹⁴, R¹⁵ and R¹⁶ each independently represents C1-4        alkyl, C2-4 alkenyl or C2-4 alkynyl, which are substituted with        1 to 5 groups selected from C1-4 alkyl, C2-4 alkenyl, C2-4        alkynyl, halogen, trifluoromethyl, OH, —COO—C1-4 alkyl, COOH,        oxo, C1-4 alkoxy, C6-C10 aryl, 5- to 10-membered heteroaryl and        NR¹⁷R¹⁸;    -   wherein R¹⁷ and R¹⁸ each independently represents (1)        hydrogen, (2) C1-4 alkyl, (3) C2-4 alkenyl or (4) C2-4 alkynyl;    -   t represents an integer of 0 to 6;    -   each R² may be the same or different and represents (1)        —COOH, (2) —COO—C1-4 alkyl, (3) —COO—C1-4 alkylene-C1-4        alkoxy, (4) —NH₂, (5) —NH—C1-4 alkyl, (6) —NH—C1-4 alkylene-C1-4        alkoxy, (7) —NHCO—C1-4 alkyl, (8) —NHCO—C1-4 alkylene-C1-4        alkoxy, (9) —NHCOO—C1-4 alkyl, (10) —NHCOO—C1-4 alkylene-C1-4        alkoxy, (11) —CONH₂, (12) —CONH—C1-4 alkyl, (13) —CONH—C2-4        alkylene-C1-4 alkoxy, (14) halogen, (15) —SO₂—C1-4 alkyl, (16)        oxo, (17) C1-4 alkoxy, (18) —CO—C1-4 alkyl, (19) —CO—C1-4        alkylene-C1-4 alkoxy or (20) —COO—C1-4 alkyl substituted with 1        to 5 groups selected from C1-4 alkyl, C2-4 alkenyl, C2-4        alkynyl, halogen, trifluoromethyl, OH, —COO—C1-4 alkyl, COOH,        oxo, C1-4 alkoxy, C6-C10 aryl, 5- to 10-membered heteroaryl and        NR¹⁹R²⁰;    -   wherein R¹⁹ and R²⁰ each independently represents (1)        hydrogen, (2) C1-4 alkyl, (3) C2-4 alkenyl or (4) C2-4 alkynyl;    -   m represents an integer of 0 to 6;    -   each R³ may be the same or different and represents (1)        —COO—C1-4 alkyl, (2) oxo, (3) —CO—C1-4 alkyl, (4) —CO—NH₂, (5)        —SO₂—NH₂ or (6) —SO₂—R⁶—R⁷;    -   n represents an integer of 0 to 6;    -   R⁶ represents (1) a bond or (2) NH;    -   R⁷ represents (1) C1-4 alkyl, (2) Cyc D or (3) C1-4 alkyl or Cyc        D substituted with 1 to 5 R⁸;    -   wherein Cyc D represents C3-C8 cycloalkyl, 5- to 10-membered        heterocycloalkyl, C6-C10 aryl or 5- to 10-membered heteroaryl;    -   each R⁸ may be the same or different and represents (1)        —COOH, (2) —COO—C1-4 alkyl, (3) —COO—C1-4 alkylene-C1-4        alkoxy, (4) —NH₂, (5) —NH—C1-4 alkyl, (6) —NHCO—C1-4 alkyl, (7)        —CONH₂, (8) —CONH—C1-4 alkyl (9) OH or (10) halogen;    -   R⁴ represents (1) a bond, (2) C1-4 alkylene, (3) C2-4 alkenylene        or (4) C2-4 alkynylene;    -   R⁵ represents (1) —CONH—, (2) Cyc E or (3) Cyc E substituted        with 1 to 5 R⁹;    -   wherein Cyc E represents C3-C8 cycloalkyl, 5- to 10-membered        heterocycloalkyl, C6-C10 aryl or 5- to 10-membered heteroaryl        and    -   each R⁹ may be the same or different and represents C1-4 alkyl        or halogen;    -   a salt thereof, an N-oxide thereof: a solvate thereof, or a        prodrug thereof.        (2) The compound according to (1), wherein the compound        represented by formula (I) represents a compound represented by        formula (I-A):

-   -   wherein all symbols have the same meanings as described above.        (3) The compound according to (2), wherein the compound        represented by formula (I-A) represents a compound represented        by formula (I-A-1):

-   -   wherein tA represents an integer of 0 to 5; and    -   the other symbols have the same meanings as described above.        (4) The compound according to (2), wherein the compound        represented by formula (I-A) represents a compound represented        by formula (I-A-A-1):

-   -   wherein all symbols have the same meanings as described above.        (5) The compound according to (1), wherein the compound        represented by formula (I) represents a compound represented by        formula (I-B):

-   -   wherein p represents an integer of 0 to 5; and    -   the other symbols have the same meanings as described above.        (6) The compound according to (5), wherein the compound        represented by formula (I-B) represents a compound represented        by formula (I-B-1):

-   -   wherein tB represents an integer of 0 to 5; and    -   the other symbols have the same meanings as described above.        (7) The compound according to (5), wherein the compound        represented by formula (I-B) represents a compound represented        by formula (I-B-B-1):

-   -   wherein all symbols have the same meanings as described above.        (8) The compound according to any one of (5) to (7), wherein Cyc        E represents imidazolyl.        (9) The compound according to (1), wherein the compound        represented by formula (I) represents a compound represented by        formula (I-C):

-   -   wherein all symbols have the same meanings as described above.        (10) The compound according to (9), wherein the compound        represented by formula (I-C) represents a compound represented        by formula (I-C-1):

-   -   wherein all symbols have the same meanings as described above.        (11) The compound according to (9), wherein the compound        represented by formula (I-C) represents a compound represented        by formula (I-C-C-1):

-   -   wherein all symbols have the same meanings as described above.        (12) The compound according to any one of (1) to (11), wherein        Cyc A represents C3-C6 cycloalkyl, C6-C10 aryl or 5- to        6-membered heterocycloalkyl.        (13) The compound according to any one of (1) to (12), wherein        Cyc A represents cyclohexyl, phenyl, piperidinyl or piperazinyl.        (14) The compound according to any one of (1) to (13), wherein        Cyc B represents C6-C10 aryl or 5- to 6-membered heteroaryl.        (15) The compound according to any one of (1) to (14), wherein        Cyc B represents phenyl or pyridyl.        (16) The compound according to any one of (1) to (15), wherein        Cyc C represents pyrrolidinyl, piperidinyl, piperazinyl or        morpholinyl.        (17) The compound according to any one of (1) to (16), wherein        -Cyc C—(R³)_(n) represents

-   -   wherein nB represents an integer of 0 to 5;    -   the arrow represents a binding position; and    -   the other symbols have the same meanings as described above.        (18) The compound according to (17), wherein -Cyc C—(R³)_(n)        represents

-   -   wherein the arrow represents a binding position; and    -   the other symbols have the same meanings as described above.        (19) The compound according to any one of (1), (2), (3),        (5), (6) or (9), wherein the compound is selected from the group        consisting of

-   (1)    4-[({(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,

-   (2)    4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,

-   (3)    4-({[(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,

-   (4)    (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-4-[4-(phenylsulfonyl)-1-piperazinyl]-2-pyrrolidinecarboxamide,

-   (5)    (2S,4S)—N-(1H-benzotriazol-6-yl)-1-{(2E)-3-[5-chloro-2-(1N-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,

-   (6)    4-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(cyclopropylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,

-   (7)    (2S,4S)-1-{(2E)-3-[S-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[(3S)-3-methyl-4-sulfamoyl-1-piperazinyl]-N-phenyl-2-pyrrolidinecarboxamide,

-   (8) methyl    [4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2    pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,

-   (9) methyl    [4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,

-   (10) methyl    [4-(2-{(2S,4R)-1-(4-carbamimidamidobenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (11) methyl    [4-(2-{(2S,4R)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (12) methyl    [4-(2-{(2S,4R)-1-[(4-carbamimidoyl-1-piperazinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-cloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (13)    4-[({(2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,

-   (14) methyl    [4-(4-chloro-2-{(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,

-   (15) methyl    [4-(2-{(2S,4R)-1-{[4-(aminomethyl)cyclohexyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl)}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (16) methyl    [4-(2-{(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,

-   (17) methyl    [4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl)}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (18) methyl    [4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,

-   (19) methyl    [4-(2-{(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,

-   (20) methyl    [4-(2-{(2S,4S)-1-[(l-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate    and

-   (21)    4-[({(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid.    (20) A pharmaceutical composition which comprises the compound    according to any one of (1) to (19), a salt thereof, an N-oxide    thereof, a solvate thereof, or a prodrug thereof.    (21) The pharmaceutical composition according to (20), which is a    factor XIa inhibitor.    (22) The pharmaceutical composition according to (21), which is an    agent for the treatment or prevention of a thromboembolic disease.    (23) The compound according to any one of (1) to (19) for use in    treating or preventing a thromboembolic disease.    (24) The compound for use according to (23), wherein the    thromboembolic disease is selected from the group consisting of    arterial cardiovascular thromboembolic disorders, venous    cardiovascular thromboembolic disorders, arterial cerebrovascular    thromboembolic disorders, venous cerebrovascular thromboembolic    disorders and thromboembolic disorders in the chambers of the heart    or in the peripheral circulation.    (25) The compound for use according to (24), wherein the    thromboembolic disease is selected from disseminated intravascular    coagulopathy (DIC), sepsis, angina, unstable angina, an acute    coronary syndrome, coronary artery disease, myocardial infarction,    atrial fibrillation, ischemic sudden death, transient ischemic    attack, stroke, acute stroke, atherothrombosis, atherosclerosis,    peripheral occlusive arterial disease, venous thrombosis, deep vein    thrombosis, thrombophlebitis, arterial embolism, coronary arterial    thrombosis, cerebral thrombosis, cerebral arterial thrombosis,    cerebral embolism, cardiogenic embolism, kidney embolism, portal    vein thrombosis, pulmonary embolism, pulmonary infarction, liver    embolism, mesenteric artery and/or vein embolism, occlusion of    retinal vein and/or artery, systemic embolism, antiphospholipid    antibody syndrome, thrombosis resulting from coronary artery bypass    graft surgery and thrombosis resulting from medical implants,    devices, or procedures in which blood is exposed to an artificial    surface that promotes thrombosis.    (26) A method for treating a patient suffering from or susceptible    to a thromboembolic disease, which method comprises administering to    said patient a therapeutically effective amount of a compound    according to any one of (1) to (19).    (27) Use of a compound according to any one of (1) to (19), in the    manufacture of a medicament for use in treating or preventing a    thromboembolic disease.    (28) The method or use according to (26) or (27), wherein the    thromboembolic disease is selected from the group consisting of    arterial cardiovascular thromboembolic disorders, venous    cardiovascular thromboembolic disorders, arterial cerebrovascular    thromboembolic disorders, venous cerebrovascular thromboembolic    disorders and thromboembolic disorders in the chambers of the heart    or in the peripheral circulation.    (29) The method or use according to (28), wherein the thromboembolic    disease is selected from unstable angina, an acute coronary    syndrome, atrial fibrillation, myocardial infarction, ischemic    sudden death, transient ischemic attack, stroke, atherosclerosis,    peripheral occlusive arterial disease, venous thrombosis, deep vein    thrombosis, thrombophlebitis, arterial embolism, coronary arterial    thrombosis, cerebral arterial thrombosis, cerebral embolism, kidney    embolism, pulmonary embolism, and thrombosis resulting from medical    implants, devices, or procedures in which blood is exposed to an    artificial surface that promotes thrombosis.    Definitions:

As used herein, a C1-4 alkyl group or moiety is a linear or branchedalkyl group or moiety containing from 1 to 4 carbon atoms. Examples ofC1-4 alkyl groups and moieties include methyl, ethyl, n-propyl,i-propyl, n-butyl, i-butyl, sec-butyl and t-butyl. For the avoidance ofdoubt, where two alkyl moieties are present in a group, the alkylmoieties may be the same or different.

In the present specification, a C1-4 alkoxy group or moiety is a linearor branched alkoxy group or moiety containing from 1 to 4 carbon atoms.Examples of C1-4 alkoxy groups and moieties include methoxy, ethoxy,n-propoxy, i-propoxy, n-butoxy, i-butoxy, sec-butoxy and t-butoxy. Forthe avoidance of doubt, where two alkoxy moieties are present in agroup, the alkoxy moieties may be the same or different.

In the present specification, the C2-4 alkenyl includes, for example,ethenyl, propenyl, butenyl and isomers thereof.

In the present specification, the C2-4 alkynyl includes, for example,ethynyl, propynyl, butynyl and isomers thereof.

In the present specification, the C1-4 alkylene includes linear orbranched alkylene such as methylene, ethylene, propylene, isopropylene,butylenes, and isobutylene.

In the present specification, the C2-4 alkenylene includes linear orbranched alkenylene such as vinylene, propenylene, 1- or 2-butenylene,and butadienylene.

In the present specification, the C2-4 alkynylene includes linear orbranched alkynylene such as ethynylene, 1- or 2-propynylene and 1- or2-butynylene.

In the present specification, the halogen atom includes, for example,fluorine, chlorine, bromine and iodine, and is preferably fluorine,chlorine or bromine.

Cyc A, Cyc B, Cyc C, Cyc D and Cyc E each independently represent C3-C8cycloalkyl, 5- to 10-membered heterocycloalkyl, C6-C10 aryl or 5- to10-membered heteroaryl.

“C3-C8 cycloalkyl” refers to a C3-C8 cyclic hydrocarbon. Examples ofC3-C8 cycloalkyl include cyclopropane, cyclobutane, cyclopentane,cyclohexane, cycloheptane, cyclooctane, cyclobutene, cyclopentene,cyclohexene, cycloheptene, cyclooctene, cyclobutadiene, cyclopentadiene,cyclohexadiene, cycloheptadiene, cyclooctadiene rings and the like.Moreover, the term “C3-C8 cycloalkyl” also includes “C3-C6 cycloalkyl”.The term “C3-C6 cycloalkyl” refers to a C3-C6 cyclic hydrocarbon.Examples of C3-C6 cycloalkyl include cyclopropane, cyclobutane,cyclopentane, cyclohexane, cyclobutene, cyclopentene, cyclohexene,cyclobutadiene, cyclopentadiene, cyclohexadiene rings and the like.

“5- to 10-membered heterocycloalkyl” refers to a “5- to 10-memberedmono- or bi-non-aromatic heterocyclic ring having 1 to 4 nitrogenatom(s), 1 or 2 oxygen atom(s) and/or 1 or 2 sulfur atom(s) as a heteroatom(s)”. Examples of 5- to 10-membered heterocycloalkyl includepyrazolidine, dihydropyridine, tetrahydropyridine, piperidine,dihydropyrazine, tetrahydropyrazine, piperazine, dihydropyrimidine,tetrahydropyrimidine, perhydropyrimidine, dihydropyridazine,tetrahydropyridazine, perhydropyridazine, dihydroazepine,tetrahydroazepine, perhydroazepine, dihydrodiazepine,tetrahydrodiazepine, perhydrodiazepine, dihydrofuran, tetrahydrofuran,dihydropyran, tetrahydropyran, dihydroxepine, tetrahydroxepine,perhydroxepine, dihydrothiophene, tetrahydrothiophene, dihydrothiopyran,tetrahydrothiopyran, dihydrothiepin, tetrahydrothiepin, perhydrothiepin,dihydroxazole, tetrahydroxazole (oxazolidine), dihydroisoxazole,tetrahydroisoxazole (isoxazolidine), dihydrothiazole, tetrahydrothiazole(thiazolidine), dihydroisothiazole, tetrahydroisothiazole(isothiazolidine), dihydrofurazan, tetrahydrofiurazan, dihydroxadiazole,tetrahydroxadiazole (oxadiazolidine), dihydroxazine, tetrahydroxazine,dihydroxadiazine, tetrahydroxadiazine, dihydroxazepine,tetrahydroxazepine, perhydroxazepine, dihydroxadiazepine,tetrahydroxadiazepine, perhydroxadiazepine, dihydrothiadiazole,tetrahydrothiadiazole (thiadiazolidine), dihydrothiazine,tetrahydrothiazine, dihydrothiadiazine, tetrahydrothiadiazine,dihydrothiazepine, tetrahydrothiazepine, perhydrothiazepine,dihydrothiadiazepine, tetrahydrothiadiazepine, perhydrothiadiazepine,morpholine, thiomorpholine, oxathiane, indoline, isoindoline,dihydrobenzofuran, perhydrobenzofuran, dihydroisobenzofuran,perhydroisobenzofuran, dihydrobenzothiophene, perhydrobenzothiophene,dihydroisobenzothiophene, perhydroisobenzothiophene, dihydroindazole,perhydroindazole, dihydroquinoline, tetrahydroquinoline,perhydroquinoline, dihydroisoquinoline, tetrahydroisoquinoline,perhydroisoquinoline, dihydrophthalazine, tetrahydrophthalazine,perhydrophthalazine, dihydronaphthyridine, tetrahydronaphthyridine,perhydronaphthyridine, dihydroquinoxaline, tetrahydroquinoxaline,perhydroquinoxaline, dihydroquinazoline, tetrahydroquinazoline,perhydroquinazoline, dihydrocinnoline, tetrahydrocinnoline,perhydrocinnoline, benzoxathiane, dihydrobenzoxazine,dihydrobenzothiazine, pyrazinomorpholine, dihydrobenzoxazole,perhydrobenzoxazole, dihydrobenzothiazole, perhydrobenzothiazole,dihydrobenzimidazole, perhydrobenzimidazole, dioxolane, 1,4-dioxane,dithiolane, dithiane, dioxaindane, benzodioxane, chroman,benzodithiolane, benzodithiane, 6,7-dihydro-5H-cyclopenta[b]pyrazine,5H-cyclopenta[b]pyrazine, 2,4-dihydro-1H-benzo[d][1,3]oxazine rings andthe like. Moreover, the term “5- to 10-membered heterocycloalkyl” alsoincludes “5- to 6-membered heterocycloalkyl”. The term “5- to 6-memberedheterocycloalkyl” refers to a “5- to 6-membered mono-non-aromaticheterocyclic ring having 1 to 3 nitrogen atom(s), 1 or 2 oxygen atom(s)and/or 1 or 2 sulfur atom(s) as a hetero atom(s)”. Examples of 5- to6-membered heterocycloalkyl include pyrazolidine, dihydropyridine,tetrahydropyridine, piperidine, dihydropyrazine, tetrahydropyrazine,piperazine, dihydropyrimidine, tetrahydropyrimidine, perhydropyrimidine,dihydropyridazine, tetrahydropyridazine, perhydropyridazine,dihydrofuran, tetrahydrofuran, dihydropyran, tetrahydropyran,dihydrothiophene, tetrahydrothiophene, dihydrothiopyran,tetrahydrothiopyran, dihydroxazole, tetrahydroxazole (oxazolidine),dihydroisoxazole, tetrahydroisoxazole (isoxazolidine), dihydrothiazole,tetrahydrothiazole (thiazolidine), dihydroisothiazole,tetrahydroisothiazole (isothiazolidine), dihydrofurazan,tetrahydrofurazan, dihydroxadiazole, tetrahydroxadiazole(oxadiazolidine), dihydroxazine, tetrahydroxazine, dihydroxadiazine,tetrahydroxadiazine, dihydrothiadiazole, tetrahydrothiadiazole(thiadiazolidine), dihydrothiadiazine, tetrahydrothiadiazine,morpholine, thiomorpholine, oxathiane, dioxolane, 1,4-dioxane,dithiolane, dithiane rings and the like.

“C6-C10 aryl” refers to a “C6-10 mono- or bi-aromatic carbocyclic ring”.Examples of C6-C10 aryl include benzene, azulene, naphthalene rings andthe like. Thus the C6-C10 aryl may be, for example, a phenyl ring andthe like.

“5- to 10-membered heteroaryl” refers to a “5- to 10-membered mono- orbi-aromatic heterocyclic ring having 1 to 4 nitrogen atom(s), 1 or 2oxygen atom(s) and/or 1 or 2 sulfur atom(s) as a hetero atom(s)”.Examples of 5- to 10-membered heteroaryl include pyrrole, imidazole,triazole, tetrazole, pyrazole, pyridine, pyrazine, pyrimidine,pyridazine, furan, thiophene, oxazole, isoxazole, thiazole, isothiazole,furazan, oxadiazole, thiadiazole, indole, isoindole, benzofuran,isobenzofuran, benzothiophene, isobenzothiophene, indazole, quinoline,isoquinoline, purine, phthalazine, pteridine, naphthyridine,quinoxaline, quinazoline, cinnoline, benzoxazole, benzothiazole,benzimidazole, benzofurazan, benzothiadiazole, benzotriazole,isoxazolo[4,5-d]pyridazine rings and the like. Moreover, the term “5- to10-membered heteroaryl” also includes “5- to 6-membered heteroaryl”. Theterm “5- to 6-membered heteroaryl” refers to a “5- to 6-memberedmono-aromatic heterocyclic ring having 1 to 3 nitrogen atom(s), 1 or 2oxygen atom(s) and/or 1 or 2 sulfur atom(s) as a hetero atom(s)”.Examples of 5- to 6-membered heteroaryl include pyrrole, imidazole,triazole, tetrazole, pyrazole, pyridine, pyrazine, pyrimidine,pyridazine, furan, thiophene, oxazole, isoxazole, thiazole, isothiazole,furazan, oxadiazole, thiadiazole rings and the like.

Cyc D represents C3-C8 cycloalkyl, 5- to 10-membered heterocycloalkyl,C5-C10 aryl or 5- to 10-membered heteroaryl, any of which may beoptionally substituted with 1 to 5 R⁸.

Cyc E represents C3-C8 cycloalkyl, 5- to 10-membered heterocycloalkyl,C5-C10 aryl or 5- to 10-membered heteroaryl, any of which may beoptionally substituted with 1 to 5R⁹.

The optionally substituted “C3-C8 cycloalkyl” represented by Cyc D orCyc E may be selected from any of the examples provided above for “C3-C8cycloalkyl”.

The optionally substituted “5- to 10-membered heterocycloalkyl”represented by Cyc D or Cyc E may be selected from any of the examplesprovided above for “5- to 10-membered heterocycloalkyl”.

The optionally substituted “C6-C10 aryl” represented by Cyc D or Cyc Emay be selected from any of the examples provided above for “C6-C10aryl”.

The optionally substituted “5- to 10-membered heteroaryl” represented byCyc D or Cyc E may be selected from any of the examples provided abovefor “5- to 10-membered heteroaryl”.

R¹ represents C6-C10 aryl or 5- to 10-membered heteroaryl, any of whichmay be optionally substituted with 1 to 5 groups selected from halogen,C1-4 alkyl, C1-4 alkoxy, —C1-4 alkylene-C1-4 alkoxy, CN, —COOH,—COO—C1-4 alkyl, —CO—NH₂, —OCONH₂, —OCONH—C1-4 alkyl, —CONH—C1-4 alkyl,—NHCOO—C1-4 alkyl and —NHCO—C1-4 alkyl.

The optionally substituted “C6-C10 aryl” represented by R¹ may beselected from any of the examples provided above for “C6-C10 aryl”.

The optionally substituted “5- to 10-membered heteroaryl” represented byR¹ may be selected from any of the examples provided above for “5- to10-membered heteroaryl”.

Preferably, Cyc A represents cyclohexyl, phenyl, piperidinyl,piperazinyl or indolyl, more preferably phenyl, cyclohexyl piperidinylor piperazinyl, and further more preferably phenyl, cyclohexyl orpiperidinyl.

Preferably, Cyc B represents C6-C10 aryl or 5- to 10-memberedheteroaryl, more preferably phenyl or pyridyl.

Preferably, Cyc C represents 5- to 10-membered heterocycloalkyl, morepreferably pyrrolidinyl, piperidinyl piperazinyl or morpholinyl, furthermore preferably piperidinyl or piperazinyl.

Preferably, Cyc D represents C3-C8 cycloalkyl or C6-C10 aryl, morepreferably cyclopropyl or phenyl, any of which may be optionallysubstituted as set out above.

Preferably, Cyc E represents 5- to 10-membered heteroaryl, morepreferably imidazolyl which may be optionally substituted as set outabove.

Preferably, each R¹ independently represents 5- to 10-memberedheteroaryl which may be optionally substituted as set out above,—C(═NH)NH₂, —NH—C(═NH)NH₂, C1-4 alkyl, —C1-4 alkylene-NH₂ or halogen,more preferably tetrazolyl, —C(═NH)NH₂, —NH—C(═NH)NH₂, —CH₂NH₂, methyl,chlorine or fluorine.

Preferably, t represents an integer of 0 to 2, more preferably 1 or 2.

Preferably, tA represents an integer of 0 or 1, more preferably 0.

Preferably, tB represents an integer of 0 or 1, more preferably 0.

Preferably, each R² independently represents (1) —COOH, (2) —COO—C1-4alkyl, (3) —NH₂, (4) —NHCOO—C1-4 alkyl, (5) halogen, (6) —SO₂—C1-4 alkylor (7) C1-4 alkoxy, more preferably —COOH, —COOMe, —NH₂, —NHCOOMe,chlorine, fluorine, —SO₂-Me or methoxy.

Preferably, m represents an integer of 0, 1 or 2, more preferably 1 or2.

Preferably, each R³ independently represents (1) —COO-Me, (2) oxo, (3)—CO-Me, (4) —CO—NH₂, (5) —SO₂—NH₂ or (6) —SO₂—R⁶—R⁷, more preferably—SO₂—R⁶—R⁷, wherein R⁶ is a bond or NH and R⁷ is preferably C1-4 alkylor Cyc D, wherein Cyc D is preferably as set out above.

Preferably, n represents an integer of 0 or 1, more preferably 1.

Preferably, nB represents an integer of 0 or 1, more preferably 0.

Preferably, R⁴ represents a bond or vinylene, more preferably a bond.

Preferably, R⁵ represents (1) —CONH—, (2) Cyc E or (3) Cyc E substitutedby with halogen (preferably chlorine), wherein Cyc E is preferably asset out above.

Preferably, p represents an integer of 0 or 5, more preferably 0 or 1.

In a preferred embodiment, Cyc A represents cyclohexyl, phenyl,piperidinyl, piperazinyl or indolyl, more preferably phenyl, cyclohexylor piperidinyl, t is 1 and R¹ represents —C(═NH)NH₂, —NH—C(═NH)NH₂ or—C1-4 alkylene-NH₂, or t is 2 and one R¹ represents tetrazolyl which maybe optionally substituted as set out above and the other R¹ representshalogen.

In a preferred embodiment, Cyc A represents cyclohexyl, phenyl,piperidinyl, piperazinyl or indolyl, more preferably phenyl, cyclohexylor piperidinyl, R⁴ represent a bond and t is 1 and R¹ represents—C(═NH)NH₂, —NH—C(═NH)NH₂ or —C1-4 alkylene-NH₂, or R⁴ representvinylene and t is 2 and one R¹ represents tetrazolyl which may beoptionally substituted as set out above and the other R¹ representshalogen.

In a preferred embodiment, Cyc B represents C6-C10 aryl or 5- to10-membered heteroaryl, more preferably phenyl or pyridyl, m is 1 and R²represents (1) —COOH, (2) —COO—C1-4 alkyl, (3) —NH₂, (4) —NHCOO—C1-4alkyl, (5) halogen, (6) —SO₂—C1-4 alkyl or (7) C1-4 alkoxy, morepreferably —COOH, —COOMe, —NH₂, —NHCOOMe, chlorine, fluorine, —SO₂-Me ormethoxy.

In a preferred embodiment, Cyc B represents C6-C10 aryl or 5- to10-membered heteroaryl, more preferably phenyl or pyridyl, R⁵ represents—CONH— and m is 1 and R² represents (1) —COOH, (2) —COO—C1-4 alkyl, (3)—NH₂, (4) —NHCOO—C1-4 alkyl, (5) halogen, (6) —SO₂—C1-4 alkyl or (7)C1-4 alkoxy, more preferably —COOH, —COOMe, —NH₂, —NHCOOMe, chlorine,fluorine, —SO₂-Me or methoxy, or R⁵ represents Cyc E or Cyc Esubstituted with halogen and m is 1 and R² represents (1) —COOH, (2)—COO—C1-4 alkyl, (3) —NHCOO—C1-4 alkyl, (4) halogen, (5) —SO₂—C1-4 alkylor (6) C1-4 alkoxy, more preferably —COOH, —COOMe, —NHCOOMe, chlorine,fluorine, —SO₂-Me or methoxy.

In a preferred embodiment, -Cyc C—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein n is 0, or nB is 0 and R³ represents —SO₂—NH₂, —SO₂—R⁷ or—SO₂—NH—R⁷.

In a preferred embodiment, -Cyc C—(R³), represents

wherein the arrow represents a binding position and

the other symbols have the same meanings as described above, preferablywherein nB is 0 and —SO₂R⁶—R⁷ represents —SO₂—R⁷ or —SO₂—NH—R⁷, morepreferably nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—C1-4 alkyl or—SO₂-cyclopropyl.

The above preferred embodiments of Cyc A, Cyc B and Cyc C—(R³)_(n) maybe included in preferred compound of the invention in any combination.

In one embodiment, preferred compounds of the present invention arepyrrolidine derivatives represented by formula (I-1):

wherein the other symbols have the same meanings as described above.Preferably Cyc A, Cyc B, Cyc C, R¹, t, R², m, R³ and n in the formula(I-1) are the preferred options as described above.

In one embodiment, preferred compounds of the present invention arepyrrolidine derivatives represented by formula (I-A):

wherein the other symbols have the same meanings as described above.Preferably Cyc A, Cyc B, Cyc C, R¹, t, R², m, R³ and n in the formula(I-A) are the preferred options as described above.

Preferred compounds of formula (I-A) are those in which:

Cyc A represents C3-C8 cycloalkyl or C6-C10 aryl;

Cyc B represents C6-C10 aryl or 5- to 10-membered heteroaryl;

Cyc C represents 5- to 10-membered heterocycloalkyl;

each R¹ independently represents 5- to 10-membered heteroaryl which maybe optionally substituted as set out above, —C(═NH)NH₂, —NH—C(═NH)NH₂,C1-4 alkyl, —C1-4 alkylene-NH₂ or halogen;

t represents an integer of 0, 1 or 2;

R² represents (1) —COOH, (2) —COO—C1-4 alkyl, (3) —NH₂, (4) —NHCOO—C1-4alkyl, (5) halogen, (6) —SO₂—C1-4 alkyl or (7) C1-4 alkoxy;

m represents an integer of 0, 1 or 2;

each R³ independently represents (1) —COO-Me, (2) oxo, (3) —CO-Me, (4)—CO—NH₂, (5) —SO₂—NH₂ or (6) —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NH andR⁷ is preferably C1-4 alkyl or Cyc D, wherein Cyc D is preferably as setout above;

n represents an integer of 0 or 1.

Preferred compounds of formula (I-A) include those in which:

Cyc A represents cyclohexyl, phenyl, piperidinyl or piperazinyl;

Cyc B represents phenyl or pyridyl;

Cyc C represents pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl;

each R¹ independently represents tetrazolyl, —C(═NH)NH₂, —NH—C(═NH)NH₂,—CH₂NH₂, methyl, chlorine or fluorine;

t represents an integer of 1 or 2;

R² represents —COOH, —COOMe, —NH₂, —NHCOOMe, chlorine, fluorine, —SO₂-Meor methoxy;

m represents an integer of 1 or 2;

each R³ independently represents —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NHand R⁷ is preferably C1-4 alkyl, cyclopropyl or phenyl;

n represents an integer of 1.

Further preferred compounds of formula (I-A) include those in which CycA represents cyclohexyl, phenyl, piperidinyl, piperazinyl or indolyl,more preferably phenyl, cyclohexyl or piperidinyl and t is 1 and R¹represents —C(═NH)NH₂, —NH—C(═NH)NH₂ or —CH₂NH₂.

Further preferred compounds of formula (I-A) include those in which CycB represents C6-C10 aryl or 5- to 10-membered heteroaryl, morepreferably phenyl or pyridyl, and m is 1 and R² represents (1) —COOH,(2) —COO—C1-4 alkyl, (3) —NHCOO—C1-4 alkyl, (4) halogen, (5) —SO₂—C1-4alkyl or (6) C1-4 alkoxy, more preferably —COOH, —COOMe, —NHCOOMe,chlorine, fluorine, —SO₂-Me.

Further preferred compounds of formula (I-A) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein n is 0, or nB is 0 and R³ represents —SO₂—NH₂, —SO₂—R⁷ or—SO₂—NH—R⁷.

Further preferred compounds of formula (I-A) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—R⁷ or —SO₂—NH—R⁷, morepreferably nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—C1-4 alkyl or—SO₂-cyclopropyl.

Further preferred compounds of formula (I-A) include a compound of(I-A-1):

wherein tA represents an integer of 0 or 1, more preferably 0, and theother symbols have the same meanings as described above and the samepreferred definitions as set out above (alone or in any combination), acompound of (I-A-2):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-3):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-4):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-5):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-6):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (1-A-7):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-8):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-9):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-10):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-A-11):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), and the like.

In another embodiment, preferred compounds of the present invention arepyridinone derivatives represented by formula (I-B):

wherein the other symbols have the same meanings as described above.Preferably, Cyc A, Cyc B, Cyc C, Cyc E, R¹, t, R², m, R³, n, R⁹ and p inthe formula (I-B) are the preferred options as described above.

Preferred compounds of formula (I-B) are those in which:

Cyc A represents C3-C8 cycloalkyl or C6-C10 aryl;

Cyc B represents C6-C10 aryl or 5- to 10-membered heteroaryl;

Cyc C represents 5- to 10-membered heterocycloalkyl;

Cyc E represents 5- to 10-membered heteroaryl;

each R¹ independently represents 5- to 10-membered heteroaryl which maybe optionally substituted as set out above, —C(═NH)NH₂, —NH—C(═NH)NH₂,C1-4 alkyl, —C1-4 alkylene-NH₂ or halogen;

t represents an integer of 0, 1 or 2;

each R² represents (1) —COOH, (2) —COO—C1-4 alkyl, (3) —NH₂, (4)—NHCOO—C1-4 alkyl, (5) halogen, (6) —SO₂—C1-4 alkyl or (7) C1-4 alkoxy;m represents an integer of 0, 1 or 2;

each R¹ independently represents (1) —COO-Me, (2) oxo, (3) —CO-Me, (4)—CO—NH₂, (5) —SO₂—NH₂ or (6) —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NH andR⁷ is preferably C1-4 alkyl or Cyc D, wherein Cyc D is preferably as setout above;

n represents an integer of 0 or 1;

each R⁹ represents halogen;

p represents an integer of 0 or 1.

Preferred compounds of formula (I-B) include those in which:

Cyc A represents cyclohexyl, phenyl, piperidinyl or piperazinyl;

Cyc B represents phenyl or pyridyl;

Cyc C represents pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl;

Cyc E represents imidazolyl;

each R¹ independently represents tetrazolyl, —C(═NH)NH₂, —NH—C(═NH)NH₂,—CH₂NH₂, methyl, chlorine or fluorine;

t represents an integer of 1 or 2;

R² represents —COOH, —COOMe, —NH₂, —NHCOOMe, chlorine, fluorine, —SO₂-Meor methoxy;

m represents an integer of 1 or 2;

each R³ independently represents —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NHand R⁷ is preferably C1-4 alkyl, cyclopropyl or phenyl;

n represents an integer of 1;

each R⁹ represents chlorine;

p represents an integer of 0 or 1.

Further preferred compounds of formula (I-B) include those in which CycA represents cyclohexyl, phenyl, piperidinyl, piperazinyl or indolyl,more preferably phenyl, cyclohexyl or piperidinyl and t is 1 and R¹represents —C(═NH)NH₂, —NH—C(═NH)NH₂, or —CH₂NH₂.

Further preferred compounds of formula (I-B) include those in which CycB represents C6-C10 aryl or 5- to 10-membered heteroaryl, morepreferably phenyl or pyridyl, and m is 1 and R² represents (1) —COOH,(2) —COO—C1-4 alkyl, (3) —NH₂, (4) —NHCOO—C1-4 alkyl, (5) halogen, (6)—SO₂—C1-4 alkyl or (7) C1-4 alkoxy, more preferably —COOH, —COOMe, —NH₂,—NHCOOMe, chlorine, fluorine, —SO₂-Me.

Further preferred compounds of formula (I-B) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein n is 0, or nB is 0 and R³ represents —SO₂—NH₂, —SO₂—R⁷ or—SO₂—NH—R⁷.

Further preferred compounds of formula (I-B) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—R⁷ or —SO₂—NH—R⁷, morepreferably nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—C1-4 alkyl or—SO₂-cyclopropyl

Further preferred compounds of formula (I-B) include a compound of(I-B-1):

wherein tB represents an integer of 0 or 1, more preferably 0, pBrepresents an integer of 0 or 1, more preferably 0, and the othersymbols have the same meanings as described above and the same preferreddefinitions as set out above (alone or in any combination), a compoundof (I-B-2):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-3):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-4):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-5):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-6):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-7):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-8):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-9):

wherein mB represents an integer of 0 to 4, more preferably 0 or 1, andthe other symbols have the same meanings as described above and the samepreferred definitions as set out above (alone or in any combination), acompound of (I-B-10):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-11):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-12):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-B-13):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), and the like.

In another embodiment, preferred compounds of the present invention arepyridinone derivatives represented by formula (I-C):

wherein the other symbols have the same meanings as described above.Preferably, Cyc A, Cyc B, Cyc C, R¹, t, R², m, R³, n and R⁵ in theformula (I-C) are the preferred options as described above.

Preferred compounds of formula (I-C) are those in which:

Cyc A represents C3-C8 cycloalkyl or C6-C10 aryl;

Cyc B represents C6-C10 aryl or 5- to 10-membered heteroaryl; Cyc Crepresents 5- to 10-membered heterocycloalkyl;

each R¹ independently represents 5- to 10-membered heteroaryl which maybe optionally substituted as set out above, —C(═NH)NH₂, —NH—C(═NH)NH₂,C1-4 alkyl, —C1-4 alkylene-NH₂ or halogen;

t represents an integer of 0, 1 or 2; each R² represents (1) —COOH, (2)—COO—C1-4 alkyl, (3) —NH₂, (4) —NHCOO—C1-4 alkyl, (5) halogen, (6)—SO₂—C1-4 alkyl or (7) C1-4 alkoxy;

m represents an integer of 0, 1 or 2;

each R³ independently represents (1) —COO-Me, (2) oxo, (3) —CO-Me, (4)—CO—NH₂, (5) —SO₂—NH₂ or (6) —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NH andR⁷ is preferably C1-4 alkyl or Cyc D, wherein Cyc D is preferably as setout above;

n represents an integer of 0 or 1;

R⁵ represents (1) —CONH—, (2) Cyc E or (3) Cyc E substituted by withhalogen.

Preferred compounds of formula (I-C) include those in which:

Cyc A represents cyclohexyl, phenyl, piperidinyl or piperazinyl;

Cyc B represents phenyl or pyridyl;

Cyc C represents pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl;

each R¹ independently represents tetrazolyl, —C(═NH)NH₂, —NH—C(═NH)NH₂,—CH₂NH₂, methyl, chlorine or fluorine;

t represents an integer of 1 or 2;

R² represents —COOH, —COOMe, —NH₂, —NHCOOMe, chlorine, fluorine, —SO₂-Meor methoxy;

m represents an integer of 1 or 2;

each R³ independently represents —SO₂—R⁶—R⁷, wherein R⁶ is a bond or NHand R⁷ is preferably C1-4 alkyl, cyclopropyl or phenyl;

n represents an integer of 1;

R⁵ represents (1) —CONH—, (2) imidazolyl or (3) imidazolyl substitutedby with chlorine.

Further preferred compounds of formula (I-C) include those in which CycA represents cyclohexyl, phenyl, piperidinyl, piperazinyl or indolyl,more preferably phenyl, t is 2 and one R¹ represents tetrazolyl whichmay be optionally substituted as set out above and the other R¹represents halogen.

Further preferred compounds of formula (I-C) include those in which CycB represents C6-C10 aryl or 5- to 10-membered heteroaryl, morepreferably phenyl, and m is 1 and R² represents (1) —COOH, (2) —COO—C1-4alkyl, (3) —NH₂, (4) —NHCOO—C1-4 alkyl, (5) halogen, (6) —SO₂—C1-4 alkylor (7) C1-4 alkoxy, more preferably —COOH, —COOMe, —NHCOOMe, chlorine,fluorine, —SO₂-Me or methoxy.

Further preferred compounds of formula (I-C) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position and

the other symbols have the same meanings as described above, preferablywherein n is 0 or nB is 0 and R³ represents —SO₂—NH₂, —SO₂—R⁷ or—SO₂—NH—R⁷.

Further preferred compounds of formula (I-C) include those in which -CycC—(R³)_(n) represents

wherein the arrow represents a binding position; and

the other symbols have the same meanings as described above, preferablywherein nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—R⁷ or —SO₂—NH—R⁷, morepreferably nB is 0 and —SO₂—R⁶—R⁷ represents —SO₂—C1-4 alkyl or—SO₂-cyclopropyl.

Further preferred compounds of formula (I-C) include a compound of(I-C-1):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-C-2):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-C-3):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), a compound of (I-C-4):

wherein all symbols have the same meanings as described above and thesame preferred definitions as set out above (alone or in anycombination), and the like.

As used herein, general references to “compounds of formula (I)” includecompounds of formula (I-A), (I-B) and (I-C).

Particularly preferred compounds of formula (I-A) include:

-   4-[({(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-(4-carbamimidamidobenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-{[cis-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-[(trans-4-carbamimidoylcyclohexyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   4-({[(3′S,5′S)-1′-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-1,3′-bipyrrolidin-5′-yl]carbonyl}amino)benzoic    acid,-   4-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(1-piperidinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   4-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methoxycarbonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-({[(2S,4S)-4-(4-acetyl-1-piperazinyl)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   4-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(4-carbamoyl-1-piperazinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   4-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(3-oxo-1-piperazinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   (2S,4S)-1-[(3-chloro-1H-indol-5-yl)carbonyl]-4-[4-(cyclopropylsulfonyl)-1-piperazinyl]-N-phenyl-2-pyrrolidinecarboxamide,-   4-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(cyclopropylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-{[4-(aminomethyl)-1-piperidinyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-[(4-carbamimidoyl-1-piperazinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(ethylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   4-[({(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   benzyl    4-[({(2S,4S)-1-(4-{N-[(benzyloxy)carbonyl]carbamimidoyl}benzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate,-   ethyl    4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate,-   benzyl    4-[({(2S,4S)-1-(4-cyanobenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate    and-   benzyl    4-[({(2S,4S)-1-[4-(N′-hydroxycarbamimidoyl)benzoyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

salts thereof, N-oxides thereof, solvates thereof, and prodrugs thereof.

Particularly preferred compounds of formula (I-B) include:

-   methyl    [4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-{[cis-4-(anilinomethyl)cyclohexyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-[(4-carbamimidoyl-1-piperazinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-(4-carbamimidamidobenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-(4-carbamimidamidobenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(4-chloro-2-{(2S,4R)-1-({4-[(methylamino)methyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   (3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)[(2S,4S)-2-(4-chloro-5-phenyl-1H-imidazol-2-yl)-4-(4-morpholinyl)-1-pyrrolidinyl]methanone,-   methyl    [4-(4-chloro-2-{(2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1    l-imidazol-5-yl)phenyl]carbamate,-   methyl    [4-(2-{(2S,4R)-1-{[4-(aminomethyl)cyclohexyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-1-[(2S,4S)-4-(4-morpholinyl)-2-(5-phenyl-1H-imidazol-2-yl)-1-pyrrolidinyl]-2-propen-1-one,-   methyl    [4-(2-{(2S,4R)-1-{[4-(aminomethyl)-1-piperidinyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate,-   (2E)-1-[(2S,4S)-2-(4-chloro-5-phenyl-1H-imidazol-2-yl)-4-(4-morpholinyl)-1-pyrrolidinyl]-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propen-1-one    and-   4-[(2S,4R)-2-[5-(6-amino-3-pyridyl)-1H-imidazol-2-yl]-4-(1-methylsulfonyl-4-piperidyl)pyrrolidine-1-carbonyl]piperidine-1-carboxamidine,

salts thereof, N-oxides thereof, solvates thereof, and prodrugs thereof.

Particularly preferred compounds of formula (I-C) include:

-   4-[({(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoic    acid,-   methyl    [4-(4-chloro-2-{(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate,-   methyl    4-({[(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoate,-   4-({[(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoic    acid,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-4-(4-sulfamoyl-1-piperazinyl)-2-pyrrolidinecarboxamide,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl})-N-phenyl-4-[4-(phenylsulfonyl)-1-piperazinyl]-2-pyrrolidinecarboxamide,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-[4-(methylsulfonyl)phenyl]-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-N-(3-pyridinyl)-2-pyrrolidinecarboxamide,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl})-4-[(3S)-3-methyl-4-sulfamoyl-1-piperazinyl]-N-phenyl-2-pyrrolidinecarboxamide,-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-(4-methoxyphenyl)-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,-   (3R,3′S,5′S)-1′-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-3-(sulfamoylamino)-1,3′-bipyrrolidine-5′-carboxamide,-   (2S,4S)—N-(1H-benzotriazol-6-yl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,-   (2S,4S)—N-(3-chlorophenyl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,-   (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-1-[(2S,4S)-4-(4-morpholinyl)-2-(5-phenyl-1H-imidazol-2-yl)-1-pyrrolidinyl]-2-propen-1-one,-   (2E)-1-[(2S,4S)-2-(4-chloro-5-phenyl-1H-imidazol-2-yl)-4-(4-morpholinyl)-1-pyrrolidinyl]-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propen-1-one,-   (2S,4S)—N-(3-chloro-4-fluorophenyl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide,    and-   (2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-N-(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl)-2-pyrrolidinecarboxamide,

salts thereof, N-oxides thereof, solvates thereof, and prodrugs thereof.

Compounds of the present invention containing one or more chiral centresmay be used in enantiomerically or diastereoisomerically pure form, orin the form of a mixture of isomers. For the avoidance of doubt, thecompounds of the invention may be used in any tautomeric form.

Unless otherwise specifically mentioned, all isomers are included in thepresent invention. For example, alkyl, alkenyl, alkynyl, alkoxy andalkylthio may be straight chain or branched. Moreover, all isomers dueto double bond, ring and fused ring (E-, Z-, cis- and trans-forms),isomers due to the presence of asymmetric carbon(s) etc. (R-, S-, α- andβ-configuration, enantiomer and diastereomer), optically activesubstances having optical rotation (D-, L-, d- and 1-forms), polarcompounds by chromatographic separation (more polar compounds and lesspolar compounds), equilibrium compounds, rotational isomers, a mixturethereof in any proportion and a racemic mixture are included in thepresent invention.

According to the present invention, symbol

represents α-configuration, symbol

represents β-configuration and symbol

presents α-configuration, β-configuration or a mixture of them. There isno particular limitation for the ratio of α-configuration andβ-configuration in the mixture.

Salts:

The salt of the compound of formula (I) includes all nontoxic salts orpharmaceutically acceptable salts. With regard to the pharmaceuticallyacceptable salts, those which are low-toxicity and soluble in water arepreferred. Examples of appropriate salts of the compound of formula (I)are salt with alkaline metal (such as potassium, sodium and lithium),salt with alkaline earth metal (such as calcium and magnesium), ammoniumsalt (such as ammonium salt, tetramethylammonium salt andtetrabutylammonium salt), salt with organic amine (such astriethylamine, methylamine, dimethylamine, cyclopentylamine,benzylamine, phenethylamine, piperidine, monoethanolamine,diethanolamine, tris(hydroxymethyl) methylamine, lysine, arginine andN-methyl-D-glucamine) and acid addition salt (such as inorganic acidsalt (e.g. hydrochloride, hydrobromide, hydroiodide, sulfate, phosphateand nitrate) and organic acid salt (e.g. formate, acetate,trifluoroacetate, lactate, tartrate, oxalate, fumarate, maleate,benzoate, citrate, methanesulfonate, ethanesulfonate, benzenesulfonate,toluenesulfonate, isothionate, glucuronate and gluconate), etc.). Thesalt of the compound of the present invention also includes solvates andalso solvates with the above-mentioned alkaline (earth) metal salt,ammonium salt, organic amine salt and acid addition salt. The solvate ispreferably low-toxic and water-soluble. Examples of an appropriatesolvate are solvates with water and with alcoholic solvent (such asethanol). The compounds of the present invention are converted tolow-toxicity salts or pharmaceutically acceptable salts by knownmethods.

Moreover, the salt includes a quaternary ammonium salt. The quaternaryammonium salt of the compound represented by formula (I) is the compoundwhere nitrogen of the compounds represented by formula (I) isquarternalized by R⁰ (R⁰ is C1-8 alkyl or C1-8 alkyl substituted byphenyl).

The salt also includes an N-oxide. The compound of the present inventioncan be converted into an N-oxide by known methods. The N-oxide is thecompound where nitrogen of the compound represented by formula (I) isoxidized.

Prodrugs:

A prodrug of the compound of formula (I) means a compound which isconverted to the compound of formula (I) by reaction with an enzyme,gastric acid or the like in the living body. For example, with regard toa prodrug of the compound of formula (I), when the compound of formula(I) has an amino group, compounds in which the amino group is, forexample, acylated, alkylated or phosphorylated (e.g. compounds in whichthe amino group of the compound of formula (I) is eicosanoylated,alanylated, pentylaminocarbonylated,(5-methyl-2-oxo-1,3-dioxolen-4-yl)methoxycarbonylated,tetrahydrofuranylated, pyrrolidylmethylated, pivaloyloxymethylated,acetoxymethylated, tert-butylated, etc.); when the compound of formula(I) has a hydroxyl group, compounds where the hydroxyl group is, forexample, acylated, alkylated, phosphorylated or borated (e.g. compoundsin which the hydroxyl group of the compound of formula (I) isacetylated, palmitoylated, propanoylated, pivaloylated, succinylated,fumarylated, alanylated or dimethylaminomethylcarbonylated); when thecompound of formula (I) has an amidino group, compounds in which theamidino group is, for example, hydroxylated, etherified or carbamated(e.g. compounds in which the amidino group of the compound of formula(I) is N,N′-dihydroxylated, N-methoxycarbonylated,N-2,2,2-trichloroethoxycarbonylated, N-ethylthiocarbonylated,N-benzyloxycarbonylated,N-(5-methyl-2-oxo-1,3-dioxol-4-en-1-yl)-methoxycarbonylated,N-phenoxycarbonylated, N-(4-fluorophenoxy)carbonylated,N-(4-methoxyphenoxy)carbonylated, 1-acetoxyethoxycarbonylated,N-ethoxycarbonyloxylated, etc.); and when the compound of formula (I)has a carboxyl group, compounds in which the carboxyl group is, forexample, esterified or amidated (e.g. compounds in which the carboxylgroup of the compound of formula (I) is converted into ethyl ester,phenyl ester, phenylethyl ester, carboxymethyl ester,dimethylaminomethyl ester, pivaloyloxymethyl ester,ethoxycarbonyloxyethyl ester, phthalidyl ester,(5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl ester,cyclohexyloxycarbonylethyl ester or methylamide). Those compounds may beproduced by a known method per se. The prodrug of the compound offormula (I) may be either a hydrate or a non-hydrate. A prodrug of thecompound of formula (I) may also be a compound which is converted to thecompound of formula (I) under physiological conditions as described in“Iyakuhin no kaihatsu, Vol. 7 (Bunshi-sekkei), pp. 163-198(Hirokawa-Shoten), 1990”. Further, the compound of formula (I) may alsobe labeled by a radio isotope (such as ²H, 3H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N,¹⁵O, ¹⁷O, ¹⁸O, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, ¹²⁵I, etc.).

Processes for the Preparation of the Compound of the Present Invention:

The compounds of the invention can, for example, be prepared accordingto the following reaction schemes.

The compound of the present invention represented by the formula (I) maybe prepared by known methods, for example, a method combining thefollowing methods, the method according to these methods, the methodsdescribed in the examples and/or methods described in ComprehensiveOrganic Transformations: A Guide to Functional Group Preparations, 2ndEdition (Richard C. Larock, John Wiley & Sons Inc, 1999), etc., whichare appropriately modified in each following method for the preparation.Salts of the starting materials may be used.

It will also be recognized that another major consideration in theplanning of any synthetic route in this field is the judicious choice ofthe protecting group used for protection of the reactive functionalgroups present in the compounds described in this invention. Protectionreactions may be carried out by the methods, for example, described inT. W. Greene, Protective Groups in Organic Synthesis, Wiley, New York,1999.

The compound of formula (I) can be prepared from a compound representedby formula (II):

wherein R¹⁻¹ and R⁴⁻¹ have the same meanings as R¹ and R⁴ respectively.When additional carboxyl or amino groups are present they are protected,if the protection is necessary, during the amidation process with acompound represented by the formula (III):

wherein R²⁻¹, R³⁻¹ and R⁵⁻¹ have the same meanings as R², R³ and R⁵respectively. When additional carboxyl, hydroxy or amino groups arepresent they are protected if protection is necessary.

The amidation reaction is well known. For example, the reaction of thecompound represented by formula (II) with the compound represented byformula (III) wherein all symbols have the same meaning described aboveis exemplified by:

(1) A reaction procedure with use of an acid halide,

(2) A reaction procedure with use of a mixed acid anhydride, and

(3) A reaction procedure with use of a condensing agent.

Referring specifically to these reaction procedures,

-   (1) The reaction procedure employing an acid halide is conducted in    practice, for example, by reacting a carboxylic acid with an acid    halogenating agent (e.g. oxalyl chloride, thionyl chloride, etc.) in    an organic solvent (e.g. chloroform, dichloromethane, diethyl ether,    tetrahydrofuran, dimethoxyethane, etc.) at a temperature from about    −20° C. to the refluxing temperature, followed by reaction of the    resultant acid halide with an amine in an organic solvent (e.g.    chloroform, dichloromethane, diethyl ether, tetrahydrofuran,    acetonitrile, ethyl acetate, etc.) or solvent-free in the presence    of a base (e.g. pyridine, triethylamine, dimethylaniline,    4-dimethylaminopyridine, diisopropylethylamine, etc.) at a    temperature of approximately 0 to 40° C. Alternatively, the    procedure can be carried out by reacting the resultant acid halide    with an amine in an organic solvent (e.g. 1,4-dioxane,    tetrahydrofuran, dichloromethane, etc.) in the presence or absence    of a phase-transfer catalyst (e.g. tetrabutylammonium chloride,    triethylbenzylammonium chloride, tri-n-octylmethylammonium chloride,    trimethyldecylammonium chloride, tetramethylammonium chloride,    trimethyldecylammonium chloride, tetramethylammonium chloride, etc.)    at a temperature of about 0 to 40° C., whilst using an aqueous    alkali solution (e.g. an aqueous sodium bicarbonate or sodium    hydroxide solution, etc.).-   (2) The reaction procedure employing a mixed acid anhydride is    conducted in practice, for example, by reacting a carboxylic acid    with an acid halide (e.g. pivaloyl chloride, tosyl chloride, mesyl    chloride, etc.) or an acid derivative (e.g. ethyl chloroformate,    isobutyl chloroformate, etc.) in an organic solvent (e.g.    chloroform, dichloromethane, diethyl ether, tetrahydrofuran, etc.)    or solvent free in the presence of base (e.g. pyridine,    triethylamine, dimethylaniline, 4-dimethylaminopyridine,    diisopropylethylamine, etc.) at a temperature of about 0 to 40° C.,    followed by reaction of the resultant mixed acid anhydride with an    amine in an organic solvent (e.g. chloroform, dichloroethane,    diethyl ether, tetrahydrofuran, etc.) at a temperature of about 0 to    40° C.-   (3) The reaction procedure with use of a condensing agent is carried    out, for example, by reacting a carboxylic acid with an amine in an    organic solvent (e.g. chloroform, dichloromethane,    N,N-dimethylformamide, diethyl ether, tetrahydrofuran, etc.) or    solvent-free in the presence or absence of a base (e.g. pyridine,    triethylamine, diisopropylethylamine, dimethylaniline,    4-dimethylaminopyridine, etc.), with use of a condensing agent (e.g.    1,3-dicyclohexylcarbodiimide (DCC),    1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC),    1,1′-carbonyldiimidazole (CDI), 2-chloro-1-methylpyridinium iodide,    1,1′-propylphosphonic acid anhydride (1-propanephosphonic acid    cyclic anhydride, PPA),    2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium    hexafluorophosphate (HATU), etc.) and with or without use of    1-hydroxybenzotriazole (HOBt), at a temperature of about 0 to 40° C.

In the course of the synthesis of the compound of the present inventionrepresented by the formula (I), the deprotection reaction can be carriedout at an appropriate synthetic stage when protective groups ofcarboxyl, hydroxy or amino groups are present.

The deprotection reactions for protective groups of carboxyl, hydroxy oramino groups are well-known and include, for example,

(1) a deprotection reaction by alkali hydrolysis,

(2) a deprotection under acidic conditions,

(3) a deprotection reaction by hydrogenolysis,

(4) a deprotection reaction of a silyl group,

(5) a deprotection reaction using a metal,

(6) a deprotection reaction using a metal complex, etc.

To explain these methods in detail:

(1) The deprotection reaction by alkali hydrolysis is carried out, forexample, in an organic solvent (methanol, tetrahydrofuran, 1,4-dioxane,etc.) using a hydroxide of alkali metals (sodium hydroxide, potassiumhydroxide, lithium hydroxide, etc.), hydroxide of alkaline earth metals(barium hydroxide, calcium hydroxide, etc.), carbonate (sodiumcarbonate, potassium carbonate, etc.) or a solution thereof or a mixturethereof at a temperature of 0 to 40° C.(2) The deprotection reaction under acidic conditions is carried out,for example, in an organic solvent (dichloromethane, chloroform,1,4-dioxane, ethyl acetate, anisole, etc.), using an organic acid(acetic acid, trifluoroacetic acid, methanesulfonic acid,p-toluenesulfonic acid, etc.) or an inorganic acid (hydrochloric acid,sulfuric acid, etc.) or a mixture thereof (hydrobromic acid/acetic acid,etc.) in the presence or absence of 2,2,2-trifluoroethanol at atemperature of 0 to 100° C.(3) The deprotection reaction by hydrogenolysis is, for example, carriedout in a solvent (e.g. ethers such as tetrahydrofuran, 1,4-dioxane,dimethoxyethane, diethyl ether, etc.; alcohols such as methanol,ethanol, etc.; benzenes such as benzene, toluene, etc.; ketones such asacetone, methyl ethyl ketone, etc.; nitriles such as acetonitrile etc.;amides such as N,N-dimethylformamide, N,N-dimethylacetamide etc.; water,ethyl acetate, acetic acid or a mixture of two or more thereof, etc.) inthe presence of a catalyst (palladium-carbon, palladium black, palladiumhydroxide, platinum oxide, Raney nickel, etc.) under an atmosphere ofhydrogen at normal or increased pressure, or in the presence of ammoniumformate at a temperature of 0 to 200° C.(4) The deprotection reaction of a silyl group is, for example, carriedout in a water-miscible organic solvent (tetrahydrofuran, acetonitrile,etc.) using tetrabutylammonium fluoride at a temperature of 0 to 40° C.(5) The deprotection reaction using a metal is carried out, for example,in an acidic solvent (acetic acid, a buffer of pH 4.2 to 7.2 or amixture of the solution thereof and an organic solvent such astetrahydrofuran etc.) in the presence of zinc powder at a temperature of0 to 40° C. optionally under sonication.(6) The deprotection reaction using a metal complex is carried out, forexample, in an organic solvent (dichloromethane, N,N-dimethylformamide,tetrahydrofuran, ethyl acetate, acetonitrile, 1,4-dioxane, ethanol,etc.), water or a mixture thereof, in the presence of a trapping reagent(tributyltin hydride, triethylsilane, dimedone, morpholine,diethylamine, pyrrolidine, 1,3-dimethylbarbituric acid, etc.), anorganic acid (acetic acid, formic acid, 2-ethylhexanecarboxylic acid,etc.) and/or a salt of an organic acid (sodium 2-ethylhexanoate,potassium 2-ethylhexanoate, etc.) in the presence or absence of aphosphine reagent (triphenylphosphine etc.) using a metal complex(tetrakis(triphenylphosphine)palladium (0), palladium(II)bis(triphenylphosphine) dichloride, palladium(II) acetate, rhodium(I)tris(triphenylphosphine) chloride, etc.) at a temperature of 0 to 40° C.

In addition to the above, deprotection reactions may be carried out bythe methods, for example, described in T. W. Greene, Protective Groupsin Organic Synthesis, Wiley, New York, 1999.

A protective group for carboxyl includes, for example, methyl, ethyl,allyl, tert-butyl, trichloroethyl, benzyl (Bn), phenacyl,p-methoxybenzyl, trityl, 2-chlorotrityl or a solid carrier containingthese structures, etc.

A protective group for hydroxy includes, for example, methyl (Me),trityl (Tr), methoxymethyl (MOM), 1-ethoxyethyl (EE),methoxyethoxymethyl (MEM), 2-tetrahydropyranyl (THP), trimethylsilyl(TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS),tert-butyldiphenylsilyl (TBDPS), acetyl (Ac), pivaloyl (Pv), benzoyl(Bz), benzyl (Bn), p-methoxybenzyl (PMB), allyloxycarbonyl (Alloc) or2,2,2-trichloroethoxycarbonyl (Troc), etc.

A protective group for amino includes, for example, benzyloxycarbonyl,tert-butoxycarbonyl, allyloxycarbonyl (Alloc),1-methyl-1-(4-biphenyl)ethoxycarbonyl (Bpoc), trifluoroacetyl,9-fluorenylmethoxycarbonyl (FMoc), benzyl (Bn), p-methoxybenzyl,benzyloxymethyl (BOM), 2-(trimethylsilyl)ethoxymethyl (SEM), etc.

Protective groups for carboxyl, hydroxy or amino group are not limitedto those described above, but include groups which are easily andselectively deprotected. For example, those groups described in T. W.Greene, Protective Groups in Organic Synthesis, Wiley, New York, 1999.

As is easily understood by those skilled in the art, the target compoundof the present invention may be prepared easily by selecting thesedeprotection reactions.

1) The compound of formula (III) wherein R⁵⁻¹ represents a carboxyamidewhich is attached to Cyc B at nitrogen atom, that is, a compoundrepresented by formula (III-1):

wherein all symbols have the same meaning described above, can beprepared as outlined in Reaction Scheme 1:

wherein Pg¹ represents a protective group for amino described above andthe other symbols have the same meaning described above.

In Reaction Scheme 1, the compound represented by formula (IV) and theamine compound represented by formula 1a can be condensed to produce thecompound represented by formula 1b by an amidation reaction as describedabove. The compound represented by formula 1b can be converted to theamine compound represented by the formula (III-1) by a deprotectionreaction as described above.

2) The compound of formula (III) wherein R⁵⁻¹ represents an imidazolewhich is attached to Cyc B at the 4-position, that is, a compoundrepresented by formula (III-2):

wherein R⁹⁻¹ have the same meanings as R⁹, and the other symbols havethe same meaning described above, can be prepared as outlined inReaction Scheme 2:

wherein X represents fluorine, chlorine, bromine or iodine, and theother symbols have the same meaning described above.

In Reaction Scheme 2, the reaction from the compound represented byformula (IV) to the compound represented by formula 2b is an alkylationreaction.

The alkylation reaction is well known. For example, the alkylationreaction of the compound represented by formula (IV) with the compoundrepresented by formula 2a can be conducted in a solvent such asN,N-dimethylformamide, tetrahydrofuran, dichloromethane, acetone oracetonitrile in the presence of a base such as sodium carbonate,potassium carbonate, cesium carbonate, sodium bicarbonate, potassiumbicarbonate, N,N-diisopropylethylamine or triethylamine at −20° C. toreflux temperature to form a compound represented by formula 2b whereinall symbols have the same meaning described above.

The reaction from the compound represented by formula 2b to the compoundrepresented by formula 2c is an imidazole formation reaction.

The imidazole formation reaction is well known. For example, thecompound represented by formula 2b can be converted to compounds offormula 2c by heating and/or microwave irradiation in the presence ofammonium acetate or ammonium trifluoroacetate in a suitable solvent suchas xylene, toluene or acetic acid.

Alternatively, the compound represented by formula 2c can be preparedfrom the compound represented by formula 2e. The reaction from thecompound represented by formula (IV) to the compound represented byformula 2e is an amidation reaction.

The amidation reaction of the compound represented by formula (IV) withthe compound represented by formula 2d can be conducted by the method asdescribed above.

The reaction from the compound represented by formula 2e to the compoundrepresented by formula 2c is an imidazole formation reaction. Theimidazole formation reaction can be carried out by the same method asdescribed above.

The compound represented by formula 2c can be converted to the aminecompound represented by the formula (III-2) by a deprotection reactionas described above.

3) The compound of formula (III) wherein R⁵ represents an imidazole ringwhich is attached to Cyc B at the 4-position and possesses R^(9-hal),that is, a compound represented by formula (III-3):

wherein R^(9-hal) represents fluorine, chlorine, bromine or iodine, andthe other symbols have the same meaning described above, can be preparedas outlined in Reaction Scheme 3.

wherein all symbols have the same meanings as described above.

In Reaction Scheme 3, the reaction from the compound represented byformula 3a to the compound represented by formula 3b is a halogenationreaction.

The halogenation reaction is well known. For example, the reaction ofthe compound represented by formula 3a with brominating or chlorinatingagent, such as N-bromosuccinimide, N-chlorosuccinimide or1,3-dichloro-5,5-dimethylhydantoin in a suitable solvent such asacetonitrile, chloroform or tetrahydrofuran from −20° C. to therefluxing temperature provides the compound represented by formula 3b.

The compound represented by formula 3b can be converted to the aminecompound represented by the formula (III-3) by a deprotection reactionas described above.

4) The compound of formula (IV) wherein Cyc C is attached to pyrrolidinering via nitrogen atom, that is, a compound represented by formula(IV-1):

wherein all symbols have the same meanings as described above, can beprepared as outlined in Reaction Scheme 4:

wherein Pg² represents a protective group for carboxyl described aboveand Lg represents triflate, tosylate or mesylate and the other symbolshave the same meaning described above.

In Reaction Scheme 4, the reaction from the compound represented byformula 4a to the compound represented by formula 4b is a sulfonateformation reaction.

The sulfonate formation reaction is well known. For example, thetreatment of the compound represented by formula 4a with a sulfonatingreagent such as trifluoromethanesulfonic anhydride, p-toluenesulfonylchloride or methanesulfonyl chloride in a solvent such astetrahydrofuran or dichloromethane in the presence of a base such asN,N-diisopropylethylamine or triethylamine at −20° C. to refluxtemperature provides a compound represented by formula 4b.

The reaction from the compound represented by formula 4b to the compoundrepresented by formula 4d is a nucleophilic substitution reaction.

The nucleophilic substitution reaction is well known. For example, thenucleophilic substitution reaction of compound 4b with compounds offormula 4c can be conducted in a solvent such as tert-butanol orN,N-dimethylformamide in the presence of a base such asN,N-diisopropylethylamine or triethylamine at 20° C. to refluxtemperature to provide the compound represented by formula 4d.

Alternatively, the compound represented by formula 4d can be preparedfrom the compound represented by formula 4e. The reaction from thecompound represented by formula 4e to the compound represented byformula 4d is a reductive amination reaction.

The reductive amination reaction of the compound represented by formula4e with the compound represented by formula 4c can be conducted in asolvent such as methanol, tetrahydrofuran, dichloromethane,1,2-dichloroethane or acetic acid in the presence of a reductant such assodium cyanoborohydride or sodium triacetoxyborohydride at −20° C. toreflux temperature to provide the compound represented by formula 4d.

The compound represented by formula 4d can be converted to the aminecompound represented by the formula (IV-1) by a deprotection reaction asdescribed above.

5) The compound of formula (IV) wherein Cyc C is appropriatelysubstituted piperidine which is attached to pyrrolidine ring at4-position of piperidine ring, that is, a compound represented byformula (IV-2):

wherein R³⁻¹ and R³⁻² has the same meanings as R³, with the proviso thata carboxyl, hydroxyl or amino group in R³⁻¹ and R³⁻² may be protected ifnecessary, can be prepared as outlined in Reaction Scheme 5:

wherein all symbols have the same meaning described above.

In Reaction Scheme 5, the reaction from the compound represented byformula a to the compound represented by formula 5b is an enol sulfonateformation reaction.

The enol sulfonate formation reaction is well known. For example, thetreatment of the compound represented by formula 5a with a sulfonatingreagent such as trifluoromethanesulfonic anhydride,N-phenyltrifluoromethanesulfonimide,2-[N,N-bis(trifluoromethanesulfonyl)amino]pyridine, p-toluenesulfonylchloride, trifluoromethanesulfonic anhydride, p-toluenesulfonyl chlorideand methanesulfonyl chloride in a solvent such as tetrahydrofuran ordichloromethane in the presence of a base such as lithiumdiisopropylamide or sodium bis(trimethylsilyl)amide at −78° C. to 0° C.provides a compound represented by formula 5b.

Suzuki coupling reaction between a compound represented by formula 5bwith an appropriately functionalized 4-pyridineboronic acid or ester inthe presence of a base such as anhydrous cesium carbonate, cesiumfluoride, sodium carbonate or potassium phosphate in a solvent such as1,4-dioxane, N,N-dimethylformamide or dimethylsulfoxide using a catalystsuch as tetrakis(triphenylphosphine)palladium(0),1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloride,palladium(II) acetate or bis(dibenzylidenacetone)palladium(0), with orwithout a phosphine ligand such as triphenylphosphine,tri-tert-butylphosphine or 1,1′-bis(diphenylphosphino)ferrocene at atemperature from about 70° C. to the refluxing temperature provides thecompounds represented by formula 5c.

In cases where suitably substituted boronic acids or esters are notcommercially available, the 4,4,5,5-tetramethyl-[1,3,2]dioxaborolaneintermediate can be prepared from the corresponding aryl halide or aryltriflate by a palladium mediated coupling with a diboron species such asbis(pinacolato)diboron using the method of Ishiyama, T. et al. (J. Org.Chem., 1995, 60(23), 7508). Alternatively, the corresponding boronicacid can be prepared by metal-halogen exchange of the aryl/heteroarylhalide, quenching with a trialkoxyborate reagent and aqueous workup toprovide the boronic acids (Miyaura, N.; Suzuki, A. Chem. Review, 1995,95, 2457).

The hydrogenation reaction of 5c can be conducted in a solvent such asmethanol, ethanol or acetic acid in the presence of a catalyst such aspalladium-carbon, palladium black, palladium hydroxide, platinum-carbonor platinum oxide under an atmospheric or increased pressure of hydrogento give a compound represented by formula 5d.

The compound represented by formula 5d can be converted to theN-substituted compounds represented by formula 5e by an amidationreaction or sulfonamide formation reaction.

The compound of formula Se wherein R³⁻² represents acyl group can beprepared by an introduction of R³⁻² group using an amidation reaction asdescribed above.

The compound of formula 5e wherein R³⁻² represents sulfonyl group can beprepared by an introduction of R³⁻² group using a sulfonamide formationreaction.

The sulfonamide formation reaction is well known. For example, thetreatment of the compound represented by formula 5d with appropriatelysubstituted sulfonating reagent such as an alkylsulfonic anhydride,alkylsulfonyl chloride or aryl sulfonyl chloride in a solvent such astetrahydrofuran or dichloromethane in the presence of a base such asN,N-diisopropylethylamine or triethylamine at −20° C. to refluxtemperature to provide the compound represented by formula 5e.

The compound represented by formula Se can be converted to the aminecompound represented by the formula (IV-2) by a deprotection reaction asdescribed above.

The compounds of the present invention can be prepared by the reactionsor modified variants of the reactions described above.

Other starting compounds or compounds used as reagents are knowncompounds which can be prepared easily by a combination of knownmethods, for example, the methods described in Comprehensive OrganicTransformations: A Guide to Functional Group Preparations, 2nd Edition(Richard C. Larock, John Willey & Sons Inc, 1999) or Elmer J. Rauckmanet al., J. Org. Chem., 1976, 41(3), 564 etc.

In each reaction of the specification the reactions with heating, aswill be apparent to those skilled in the art, may be carried out using awater bath, an oil bath, a sand bath, a heating block or by microwave.

In each reaction of the specification, a solid phase reagent may be usedwhich is supported by a polymer (for example polystyrene,polyacrylamide, polypropylene or polyethyleneglycol etc.).

In each reaction of the specification, the products obtained may bepurified by conventional techniques. For example, the purification maybe carried out by distillation at atmospheric or reduced pressure, byhigh performance liquid chromatography with silica gel or magnesiumsilicate, by thin layer chromatography, by ion-exchange resin, byscavenger resin, by column chromatography, by washing, trituration orrecrystallization. The purification may be carried out after eachreaction stage or after several reaction stages.

In a reaction of the specification where polystyrene resin is used, theobtained products may be purified by conventional techniques. Forexample, the purification may be carried out by multiple washing with asolvent (for example, N,N-dimethylformamide, dichloromethane, methanol,tetrahydrofuran, toluene, acetic acid/toluene, etc.).

Toxicity:

The compound represented by formula (I), the salt thereof, the N-oxidethereof, the solvate thereof or the prodrug thereof show low toxicity(e.g. acute toxicity, chronic toxicity, genotoxicity, developmentaltoxicity, cardiac toxicity, drug interaction, carcinogenicity) and lackside effects such as bleeding. It may therefore be considered safe forpharmaceutical use.

Application to Pharmaceuticals:

The compounds of the present invention are therapeutically useful. Thepresent invention therefore provides a compound of formula (I), asdefined above, or a pharmaceutically acceptable salt thereof, an N-oxidethereof, a solvate thereof or a prodrug thereof, for use in thetreatment of the human or animal body by therapy.

Also provided is a pharmaceutical composition comprising a compound offormula (I), as defined above, or a pharmaceutically acceptable saltthereof, an N-oxide thereof, a solvate thereof or a prodrug thereof, anda pharmaceutically acceptable carrier or diluent.

Said pharmaceutical composition typically contains up to 85 wt % of acompound of the invention. More typically, it contains up to 50 wt % ofa compound of the invention. Preferred pharmaceutical compositions aresterile and pyrogen free. Further, the pharmaceutical compositionsprovided by the invention typically contain a compound of the inventionwhich is a substantially pure optical isomer.

The compounds of the present invention may normally be administeredsystemically or locally, usually by oral, parenteral or continuousadministration.

A therapeutically effective amount of a compound of the invention isadministered to a patient. The doses to be administered are determineddepending upon, for example, age, body weight, symptom, the desiredtherapeutic effect, the route of administration, and the duration of thetreatment. In the human adult, the doses per person are generally from 1mg to 1000 mg, by oral administration, up to several times per day, andfrom 1 mg to 100 mg, by parenteral administration (preferablyintravenous administration), up to several times per day, or continuousadministration from 1 to 24 hours per day from vein.

As mentioned above, the doses to be used depend upon various conditions.Therefore, there are cases in which doses lower than or greater than theranges specified above may be used.

The compounds or pharmaceutical compositions of the present inventionmay be administered, for example, in the form of a solid for oraladministration, liquid forms for oral administration, injections,liniments or suppositories for parenteral administration. Solid formsfor oral administration include compressed tablets, pills, capsules,dispersible powders, and granules. Capsules include hard capsules andsoft capsules.

In such solid forms, one or more of the active compound(s) may beadmixed with vehicles (such as lactose, mannitol, glucose,microcrystalline cellulose or starch), binders (such as hydroxypropylcellulose, polyvinylpyrrolidone or magnesium metasilicate aluminate),disintegrants (such as cellulose calcium glycolate), lubricants (such asmagnesium stearate), stabilizing agents, solution adjuvants (such asglutamic acid or aspartic acid, disaggregating agents, e.g. starch,alginic acid, alginates or sodium starch glycolate; effervescingmixtures, dyestuffs, sweeteners, wetting agents, such as lecithin,polysorbates, laurylsulphates; and, in general, non-toxic andpharmacologically inactive substances used in pharmaceuticalformulations and prepared according to methods well known in normalpharmaceutical practice, for example, by means of mixing, granulating,tableting, sugar coating, or film coating processes. The solid formsmay, if desired, be coated with coating agents (such as sugar, gelatin,hydroxypropyl cellulose or hydroxypropylmethyl cellulose phthalate) orbe coated with two or more films. Furthermore, coating may includecontainment within capsules of absorbable materials such as gelatin.

Liquid forms for oral administration include pharmaceutically acceptablesolutions, suspensions, emulsions, syrups and elixirs. In such forms,one or more of the active compound(s) may be dissolved, suspended oremulsified into diluent(s) commonly used in the art (such as purifiedwater, ethanol or a mixture thereof). Besides such liquid forms may alsocomprise some additives, such as wetting agents, suspending agents,emulsifying agents, sweetening agents, flavoring agents, aroma,preservative or buffering agent. The syrups may contain as carriers, forexample, saccharose or saccharose with glycerine and/or mannitol and/orsorbitol.

Suspensions and emulsions may contain a carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose or polyvinyl alcohol. The suspension or solutionsfor intramuscular injections may contain, together with the activecompound, a pharmaceutically acceptable carrier e.g. sterile water,olive oil, ethyl oleate, glycols (e.g. propylene glycol) and, ifdesired, a suitable amount of lidocaine hydrochloride.

Solutions for injection or infusion may contain as carrier, for example,sterile water or preferably they may be in the form of sterile, aqueous,isotonic saline solutions.

Injections for parenteral administration include sterile aqueoussuspensions, emulsions and solid forms which are dissolved or suspendedinto solvent(s) for injection immediately before use. In injections, oneor more of the active compound(s) may be dissolved, suspended oremulsified into solvent(s). The solvents may include distilled water forinjection, saline, vegetable oil, propylene glycol, polyethylene glycol,alcohol such as ethanol, or a mixture thereof. Injections may comprisesome additives, such as stabilizing agents, solution adjuvants (such asglutamic acid, aspartic acid or POLYSORBATE80 (registered trade mark)),suspending agents, emulsifying agents, soothing agents, buffering agentsor preservatives. They may be sterilized at a final step, or may beprepared according to sterile methods. They may also be manufactured inthe form of sterile solid forms such as freeze-dried products, which maybe dissolved in sterile water or some other sterile diluent(s) forinjection immediately before use.

Other forms for parenteral administration include liquids for externaluse, ointments and endermic liniments, inhalations, sprays,suppositories and vaginal suppositories which comprise one or more ofthe active compound(s) and may be prepared by methods known per se.

Sprays may comprise additional substances other than diluents usedcommonly, stabilizers such as sodium hydrogensulfite and buffers capableof imparting isotonicity, for example, isotonic buffers such as sodiumchloride, sodium citrate or citric acid.

Effect of the Invention

The compounds of the present invention represented by formula (I) act aspotent and selective inhibitors of factor XIa, and also show superiorproperties as a pharmaceutical product such as stability, watersolubility and the like. Thus the compounds of the present invention areuseful in preventing and/or treating thromboembolic diseases. Oneadvantage of the compounds of the present invention is that they canprovide high inhibitory activity against FXIa and high safety withoutside effects such as bleeding.

The present invention therefore provides a compound of formula (I), asdefined above, or a pharmaceutically acceptable salt thereof, an N-oxidethereof, a solvate thereof or a prodrug thereof, for use in treating orpreventing a thromboembolic disease. Also provided is a method fortreating a patient suffering from or susceptible to a thromboembolicdisease, which method comprises administering to said patient aneffective amount of a compound of formula (I), as defined above, or apharmaceutically acceptable salt thereof, an N-oxide thereof, a solvatethereof or a prodrug thereof. Further provided is the use of a compoundof formula (I), as defined above, or a pharmaceutically acceptable saltthereof, an N-oxide thereof, a solvate thereof or a prodrug thereof, inthe manufacture of a medicament for use in treating or preventing athromboembolic disease.

The thromboembolic disease may be, for example, selected from the groupconsisting of arterial cardiovascular thromboembolic disorders, venouscardiovascular thromboembolic disorders, arterial cerebrovascularthromboembolic disorders, venous cerebrovascular thromboembolicdisorders and thromboembolic disorders in the chambers of the heart orin the peripheral circulation.

More specifically, arterial cardiovascular thromboembolic disorders maybe exemplified by coronary artery disease, ischemic cardiomyopathy,acute coronary syndrome, coronary arterial thrombosis, ischemiccomplications of unstable angina and non-Q-wave myocardial infarction,acute non ST-segment elevation and/or ST-segment elevation myocardialinfarction managed medically or with subsequent percutaneous coronaryintervention, angina pectoris such as stable effort angina pectoris,variant angina pectoris, unstable angina pectoris, myocardial infarction(e.g. first myocardial infarction or recurrent myocardial infarction),acute myocardial infarction, reocclusion and restenosis after coronaryartery bypass surgery, reocclusion and restenosis after percutaneoustransluminal cardiac angioplasty/transluminal coronary artery stentplacement surgery or after thrombolytic therapy for coronary artery,ischemic sudden death. Venous cardiovascular thromboembolic disordersmay be exemplified by deep vein thrombosis (DVT) and/or pulmonaryembolism (PE) in major general surgery, abdominal surgery, hipreplacement surgery, knee replacement surgery, hip fracture surgery,multiple fracture, multiple injury, trauma, spinal cord injury, burns,critical care unit, DVT and/or PE in medical patients with severelyrestricted mobility during acute illness, DVT and/or PE in patients withcancer chemotherapy, DVT and/or PE in patients with stroke, symptomaticor asymptomatic DVT with or without PE (pulmonary embolism). Arterialcerebrovascular thromboembolic disorders may be exemplified by stroke,ischemic stroke, acute stroke, stroke in patients with non-valuvelar orvaluvelar atrial fibrillation, cerebral arterial thrombosis, cerebralinfarction, transient ischemic attack (TIA), lacuna infraction,atherosclerotic thrombotic cerebral infarction, cerebral arteryembolism, cerebral thrombosis, cerebrovascular disorder and asymptomaticcerebral infarction. Venous cerebrovascular thromboembolic disorders maybe exemplified by intracranial venous thrombosis, cerebral embolism,cerevral thrombosis, sinus thrombosis, intracranial venous sinusthrombosis and cavernous sinus thrombosis. Thromboembolic disorders inthe chambers of the heart or in the peripheral circulation may beexemplified by venous thrombosis, systemic venous thromboembolism,thrombophlebitis, non-valuvelar or valuvelar atrial fibrillation,cardiogenic embolism, disseminated intravascular coagulopathy (DIC),sepsis, acute respiratory distress syndrome (ARDS), acute lung injury(ALI), antiphospholipid antibody syndrome, kidney embolism,atherosclerosis, atherothrombosis, peripheral artery occlusive disease(PAOD), peripheral arterial disease, arterial embolism, and thrombosisresulting from medical implants, devices, or procedures in which bloodis exposed to an artificial surface (such as catheters, stents,artificial heart valves or hemodialyzer) that promotes thrombosis.

Preferably, the thromboembolic disorder is selected from unstableangina, an acute coronary syndrome, atrial fibrillation, myocardialinfarction (e.g. first myocardial infarction or recurrent myocardialinfarction), ischemic sudden death, transient ischemic attack, stroke,atherosclerosis, peripheral occlusive arterial disease, venousthrombosis, deep vein thrombosis, thrombophlebitis, arterial embolism,coronary arterial thrombosis, cerebral arterial thrombosis, cerebralembolism, kidney embolism, portal vein thrombosis, pulmonary embolism,pulmonary infarction, liver embolism, mesenteric artery and/or veinembolism, occlusion of retinal vein and/or artery, systemic embolism,disseminated intravascular coagulopathy (DIC), acute respiratorydistress syndrome (ARDS), acute lung injury (ALI), antiphospholipidantibody syndrome, thrombosis resulting from coronary artery bypassgraft surgery and thrombosis resulting from medical implants, devices,or procedures in which blood is exposed to an artificial surface (suchas catheters, stents or artificial heart valves) that promotesthrombosis.

The compounds of the present invention may also be administered incombination with one or more further therapeutic agents. Thus, inanother embodiment, the present invention provides a method for treatinga thromboembolic disorder comprising: administering to a patient in needthereof a therapeutically effective amount of a first and secondtherapeutic agent, wherein the first therapeutic agent is a compound offormula (I), as defined above, or a pharmaceutically acceptable saltthereof, an N-oxide thereof, a solvate thereof or a prodrug thereof, andthe second therapeutic agent is at least one agent selected from asecond factor XIa inhibitor, an anti-coagulant agent, an anti-plateletagent, a thrombin inhibiting agent, a thrombolytic agent, a fibrinolyticagent, a serine protease inhibitor, an elastase inhibitor and ansteroid. Preferrably, the second therapeutic agent is at least one agentselected from warfarin, unfractionated heparin, low molecular weightheparin, enoxaparin, dalteparin, bemiparin, tinzaparin, semuloparin,danaparoid, synthetic pentasaccharide, fondaparinux, hirudin,disulfatohirudin, lepirudin, bivalirudin, desirudin, argatroban,aspirin, ibuprofen, naproxen, sulindac, indomethacin, mefenamate,droxicam, diclofenac, sulfinpyrazone, piroxicam, ticlopidine,clopidogrel, prasugrel, ticagrelor, cangrelor, elinogrel, cilostazol,sarpogrelate, iroprost, beraprost, limaprost, tirofiban, eptifibatide,abciximab, melagatran, ximelagatran, dabigatran, rivaroxaban, apixaban,edoxaban, darexaban, betrixaban, TAK-442, tissue plasminogen activator,modified tissue plasminogen activator, anistreplase, urokinase,streptokinase, gabexate, gabexate mesilate, nafamostat, sivelestat,sivelestat sodium hydrate, alvelestat, ZD-8321/0892, ICI-200880,tiprelestat, elafin, alpha1-antitrypsin, cortisone, betamethasone,dexamethasone, hydrocortisone, methylprednisolone, prednisolone andtriamcinolone. Preferrably, the second therapeutic agent is at least oneanti-platelet agent. Preferrably, the anti-platelet agent(s) areclopidogrel, prasugrel, ticagrelor, cangrelor, elinogrel, cilostazol,sarpogrelate, iroprost, beraprost, limaprost and/or aspirin, or acombination thereof. The present invention also provides a compound offormula (I), as defined above, or a pharmaceutically acceptable saltthereof, an N-oxide thereof, a solvate thereof or a prodrug thereof, incombination with a second therapeutic agent selected from those listedabove, for use in treating or preventing a thromboembolic disease. Thepresent invention also provides the use of a compound of formula (I), asdefined above, or a pharmaceutically acceptable salt thereof, an N-oxidethereof, a solvate thereof or a prodrug thereof, in combination with asecond therapeutic agent, in the manufacture of a medicament for use intreating or preventing a thromboembolic disease.

In another embodiment, the present invention provides a pharmaceuticalcomposition comprising a compound of formula (I), as defined above, or apharmaceutically acceptable salt thereof, an N-oxide thereof, a solvatethereof or a prodrug thereof and an additional therapeutic agent.Preferably, the further additional therapeutic agent(s) are selectedfrom potassium channel openers, potassium channel blockers, calciumchannel blockers, sodium hydrogen exchanger inhibitors, antiarrhythmicagents, antiatherosclerotic agents, anticoagulants, antiplatelets,antithrombotic agents, prothrombolytic agents, fibrinogen antagonists,diuretics, antihypertensive agents, ATPase inhibitors, mineralcorticoidreceptor antagonists, phospodiesterase inhibitors, antidiabetic agents,protease inhibitors, elastase inhibitors, anti-inflammatory agents,antioxidants, angiogenesis modulators, antiosteoporosis agents, hormonereplacement therapies, hormone receptor modulators, oral contraceptives,antiobesity agents, antidepressants, antianxiety agents, antipsychoticagents, antiproliferative agents, antitumor agents, antiulcer andgastroesophageal reflux disease agents, growth hormone agents and/orgrowth hormone secretagogues, thyroid mimetics, anti-infective agents,antiviral agents, antibacterial agents, antifungal agents,cholesterol/lipid lowering agents and lipid profile therapies, andagents that mimic ischemic preconditioning and/or myocardial stunning,or a combination thereof,

In another embodiment, the present invention provides a pharmaceuticalcomposition further comprising additional therapeutic agent(s) selectedfrom an anti-arrhythmic agent, an anti-hypertensive agent, ananti-coagulant agent, an anti-platelet agent, a thrombin inhibitingagent, a thrombolytic agent, a fibrinolytic agent, a calcium channelblocker, a potassium channel blocker, a cholesterol/lipid loweringagent, a serine protease inhibitor, an elastase inhibitor, ananti-inflammatory agent, or a combination thereof.

In another embodiment, the present invention provides a pharmaceuticalcomposition further comprising additional therapeutic agent(s) selectedfrom warfarin, unfractionated heparin, low molecular weight heparin,enoxaparin, dalteparin, bemiparin, tinzaparin, semuloparin, danaparoid,synthetic pentasaccharide, fondaparinux, hirudin, disulfatohirudin,lepirudin, bivalirudin, desirudin, argatroban, aspirin, ibuprofen,naproxen, sulindac, indomethacin, mefenamate, dipyridamol, droxicam,diclofenac, sulfinpyrazone, piroxicam, ticlopidine, clopidogrel,prasugrel, ticagrelor, cangrelor, elinogrel, cilostazol, sarpogrelate,iroprost, beraprost, limaprost, tirofiban, eptifibatide, abciximab,melagatran, ximelagatran, dabigatran, rivaroxaban, apixaban, edoxaban,darexaban, betrixaban, TAK-442, tissue plasminogen activator, modifiedtissue plasminogen activator, anistreplase, urokinase, streptokinasegabexate, gabexate mesilate, nafamostat, sivelestat, sivelestat sodiumhydrate, alvelestat, ZD-8321/0892, ICI-200880, tiprelestat, elafin,alpha1-antitrypsin, cortisone, betamethasone, dexamethasone,hydrocortisone, methylprednisolone, prednisolone and triamcinolone or acombination thereof.

In a preferred embodiment, the present invention provides apharmaceutical composition wherein the additional therapeutic agent isan antihypertensive agent selected from ACE inhibitors, AT-1 receptorantagonists, beta-adrenergic receptor antagonists, ETA receptorantagonists, dual ETA/AT-1 receptor antagonists, and vasopepsidaseinhibitors, an antiarrythmic agent selected from IKur inhibitors,elastase inhibitors, serine protease inhibitors, steroids, ananticoagulant selected from thrombin inhibitors, antithrombin-IIIactivators, heparin co-factor II activators, other factor XIainhibitors, plasma and/or tissue kallikrein inhibitors, plasminogenactivator inhibitor (PAI-1) inhibitors, thrombin activatablefibrinolysis inhibitor (TAFI) inhibitors, factor VIIa inhibitors, factorVIIIa inhibitors, factor IXa inhibitors, factor Xa inhibitors and factorXIIa inhibitors, or an anti-platelet agent selected from GPII/IIablockers, protease activated receptor (PAR-1) antagonists, PAR-4antagonists, phosphodiesterase-III inhibitors, other phosphodiesteraseinhibitors, P2X1 antagonits, P2Y₁ receptor antagonists, P2Y₁₂antagonists, thromboxane receptor antagonists, thromboxane A2 synthaseinhibitors, cyclooxygense-1 inhibitors, phospholipase D1 inhibitors,phospholipase D2 inhibitors, phospholipase D inhibitors, glycoprotein VI(GPVI) antagonists, glycoprotein Ib (GPIb) antagonists, Growtharrest-specific gene 6 product (Gas6) antagonists and aspirin, or acombination thereof.

In a preferred embodiment, the present invention provides apharmaceutical composition, wherein the additional therapeutic agent(s)are an anti-platelet agent or a combination thereof.

Best Mode for Carrying Out the Invention

The present invention is illustrated by the following Examples andbiological Examples, but it is not limited thereto.

The solvents in the parentheses described in chromatographic separationand TLC show the eluting or developing solvents, and the ratios of thesolvents used are given as percentage mixtures in chromatographicseparations or TLC. Where a compound is described as dried, eitheranhydrous magnesium or sodium sulphate was used. The solvents in theparentheses in NMR show the solvents used in measurement. DMSO-d₆represents deuterated dimethylsulfoxide; CDCl₃ represents deuteratedchloroform; CD₃OD represents deuterated methanol; D₂O representsdeuterated water. The following abbreviations are used in reporting the¹H NMR spectra: s (singlet), d (doublet), t (triplet), q (quartet),quint. (quintet), br. (broad), app. (apparent), obs. (obscured).

Including compounds in the following Examples, compounds used in thepresent specification were commonly named using a computer programcapable of naming in accordance with IUPAC rules; ACD/Name manufacturedby Advanced Chemistry Development Inc., JChem for Excel or MarvinSketchmanufactured by ChemAxon Ltd., or IUPAC nomenclature. In each of thefollowing Examples, the name of the objective compound of the Example isdescribed subsequently to the number of the Example, and the compound issometimes referred to as the “title compound”.

Example 1 methyl (2S,4R)-4-hydroxy-2-pyrrolidinecarboxylatehydrochloride

To a solution of (2S,4R)-4-hydroxy-2-pyrrolidinecarboxylic acidhydrochloride (1.0 g, 7.6 mmol) in methanol (25 mL) at 0° C. was addedthionyl chloride (0.83 mL, 11.4 mmol). The reaction was warmed to roomtemperature and heated at reflux overnight. After cooling to roomtemperature, the reaction mixture was concentrated to dryness to givethe title compound (1.2 g, 92%) as a white solid.

¹H NMR (300 MHz, DMSO-d₆) δ 9.99 (br. s, 2H), 5.58 (br. s, 1H),4.49-4.41 (m, 2H), 3.75 (s, 3H), 3.38 (dd, 1H), 3.07 (d, 1H), 2.23-2.04(m, 2H).

Example 2 1-benzyl 2-methyl(2S,4R-4-hydroxy-1,2-pyrrolidinedicarboxylate

Chlorobenzylformate (1.0 mL, 7.9 mmol) was added to a mixture of thecompound prepared in Example 1 (1.2 g, 6.6 mmol), NaHCO₃ (4.0 g) andsaturated aqueous NaHCO₃ (5.0 mL) in THF (20 mL) at 0° C. After stirringat room temperature for 3 h, tris(hydroxymethyl)aminomethane (1.4 g) wasadded and the reaction mixture was partitioned between ethyl acetate andwater, the combined organic extracts were dried and concentrated andpurified by flash chromatography (silica gel, 40 g, 20-80% ethylacetate/hexanes) to give the title compound (1.3 g, 72%) as a paleyellow oil.

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.35-7.29 (m, 5H), 5.18-4.97(m, 2H), 4.47-4.39 (m, 2H), 3.71 (s, 1.5H), 3.62-3.51 (m, 3.5H),2.30-2.25 (m, 1H), 2.09-2.00 (m, 1H).

Example 3 1-benzyl 2-methyl(2S,4S)-4-[4-(methylsulfonyl)-1-piperazinyl]-1,2-pyrrolidinedicarboxylate

To a solution of the compound prepared in Example 2 (1.0 g, 3.6 mmol)and N,N-diisopropylethylamine (1.24 mL, 7.16 mmol) in dichloromethane(20 mL) at −20° C. was added trifluoromethylsulfonic anhydride (0.904 g,5.36 mmol). The reaction mixture was stirred at room temperature for 1 hthen concentrated to dryness to obtain the crude triflate. The crudematerial was dissolved in tert-butanol (50 mL) andN,N-diisopropylethylaminc (1.24 mL, 7.16 mmol) and1-(methylsulfonyl)piperazine (1.76 g, 10.7 mmol) were added to thereaction at room temperature. The resulting mixture was heated at 100°C. for 48 h. After cooling to room temperature, the solvent was removedand the crude reaction mixture was purified by flash chromatography(silica gel, 40 g, 20-60% ethyl acetate/hexanes) to give the titlecompound (2.35 g, 85%) as an off-white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 7.35-7.28 (m, 5H), 5.19-5.00(m, 2H), 4.15-4.31 (m, 1H), 3.98-3.82 (m, 1H), 3.75 (s, 1.7H), 3.55 (s,1.3H), 3.33-3.22 (m, 5H), 2.91-2.81 (m, 1H), 2.76 (s, 3H), 2.62-2.46 (m,5H), 1.91-1.81 (m, 1H).

Example 4(2S,4S)-1-[(benzyloxy)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinecarboxylicacid

To a solution of the compound prepared in Example 3 (0.90 g, 2.11 mmol)in tetrahydrofuran (20 mL) and water (20 mL) at 0° C. was added lithiumhydroxide (0.203 g, 8.4 mmol). The reaction was warmed to roomtemperature and stirred overnight. The reaction mixture was carefullyacidified to pH 5 with 2 M hydrochloric acid. The aqueous solution wasextracted with ethyl acetate (2×300 mL) and the combined organicextracts were dried and concentrated to give the title compound (0.565g, 65%) as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 7.36-7.33 (m, 5H), 5.18-5.12(m, 2H), 4.43-4.36 (m, 1H), 3.98-3.82 (m, 2H), 3.37-3.17 (m, 4H),3.00-2.91 (m, 1H), 2.82-2.71 (m, 6H), 2.64-2.49 (m, 2H), 2.24-2.12 (m,1H), 1.97-1.85 (m, 1H).

Example 5 benzyl(2S,4S)-2-[(4-{[(2-methyl-2-propanyl)oxy]carbonyl}phenyl)carbamoyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-1-pyrrolidinecarboxylate

To a solution of the compound prepared in Example 4 (0.20 g, 0.40 mmol)and 2-methyl-2-propanyl 4-aminobenzoate (0.154 g, 0.80 mmol) in pyridine(50 mL) was added 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimidehydrochloride (0.630 mg 3.2 mmol) at 0° C. The reaction was stirred atroom temperature for 18 h. The mixture was concentrated under reducedpressure and the resulting residue diluted with dichloromethane (20 mL).This solution was washed with brine, dried and concentrated.Purification by flash chromatography (silica gel, 40 g, 20-80% ethylacetate/hexanes) gave the title compound (0.185 g, 65%) as a whitesolid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 9.15 (br. s, 0.6H), 8.33(br. s, 0.4H), 7.88 (d, 2H), 7.50 (d, 2H), 7.35-7.01 (m, 5H), 5.22-4.93(m, 2H), 4.56-4.36 (m, 1H), 3.88-3.76 (m, 1H), 3.22-3.00 (m, 4H),2.93-2.78 (m, 3H), 2.69-2.48 (m, 6H), 2.45-2.24 (m, 2H), 1.58 (s, 9H).

Example 6 2-methyl-2-propanyl4-[({(2S,4S)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

To a solution of the compound prepared in Example 5 (2.1 g, 7.4 mmol) inethanol (100 mL) was added Pd/C (0.40 g, 20% by wt). The reaction wasstirred under an atmosphere of hydrogen (50 psi) at room temperature for6 h. The reaction mixture was filtered through diatomaceous earth andconcentrated to give the title compound (1.10 g, 69%) as a white solid.

¹H NMR (400 MHz, CDCl₃) δ 9.85 (s, 1H), 7.95 (d, 2H), 7.63 (d, 2H), 3.95(dd, 1H), 3.28 (dd, 1H), 3.14 (t, 4H), 2.89-2.85 (m, 1H), 2.83-2.76 (m,1H), 2.63 (s, 3H), 2.60-2.44 (m, 5H), 2.05 (br. s, 1H), 2.03-1.98 (m,1H), 1.58 (s, 9H).

Example 71-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid

To a solution of piperidine-4-carboxylic acid trifluoroacetate salt(0.20 g, 0.82 mmol) in methanol (10 mL), triethylamine (0.20 mL, 1.6mmol) andN,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}-1H-pyrazole-1-carboximidamide(0.30 g, 0.98 mmol) was added and the reaction mixture stirred at roomtemperature for 18 h. The solvent was evaporated under reduced pressureand the resulting residue dissolved in ethyl acetate and washed withbrine. The organic layer was dried and concentrated to dryness to obtainthe title compound (200 mg, 76%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 3.91 (d, 2H), 3.30 (t, 2H), 2.63-2.66 (m,1H), 1.90-1.96 (m, 2H), 1.67-1.69 (m, 2H), 1.45 (s, 18H).

Example 8 2-methyl-2-propanyl4-[({(2S,4S)-1-{[1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

To a solution of the compound prepared in Example 6 (0.2 g, 0.7 mmol) inN,N-dimethylformamide (2 mL) was added2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (0.10 g, 0.26 mmol) at 0° C. After stirring for 20minutes, the compound prepared in Example 7 (0.12 g, 0.26 mmol) andN,N-diisopropylethylamine (0.15 mL, 0.8 mmol) were added and thereaction stirred at room temperature for 2 h. The reaction was quenchedby adding ice cold water and the resulting precipitate was collected byfiltration, dried and the crude product purified by flash chromatographyto afford the title compound (0.11 g, 61%) as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 9.35 (s, 1H), 7.92 (d, 2H),7.52 (d, 2H), 4.75 (t, 1H), 4.28-4.10 (m, 2H), 3.88-3.84 (m, 1H), 3.37(t, 1H), 3.34 (t, 3H), 3.10-2.93 (m, 4H), 2.75 (s, 3H), 2.73-2.57 (m,6H), 2.30-2.22 (m, 1H), 1.83-1.75 (m, 4H), 1.57 (s, 9H), 1.47 (s, 18H).

Example 94-[({(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

To a solution of the compound prepared in Example 8 (0.11 g, 0.13 mmol)in dichloromethane (15 mL) at 0° C. was added trifluoroacetic acid (0.5mL). The reaction was warmed to room temperature and stirred for 18 h.The solvent and excess trifluoroacetic acid was removed under reducedpressure. The solid was dissolved in water and lyophilized to dryness toafford the title compound (0.030 g, 40%) as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) 7.97 (d, 2H), 7.69 (d, 2H),4.55 (t, 1H), 4.26-4.01 (m, 1H), 3.93-3.87 (m, 2H), 3.72 (t, 1H),3.55-3.51 (m, 1H), 3.41 (br. s, 4H), 3.25-3.04 (m, 6H), 3.05-2.91 (m,1H), 2.90 (s, 3H), 2.78-2.71 (m, 1H), 2.20-2.06 (m, 1H), 2.00-1.96 (m,1H), 1.89-1.85 (m, 1H), 1.73-1.67 (m, 2H).

ESI MS m/z 550 (M+H)⁺

Example 104-(N′,N″-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidamido)benzoicacid

To a solution of 4-carbamimidamidobenzoic acid hydrochloride (0.27 g,1.3 mmol) in ethanol (10 mL), 2 M aqueous NaOH (1.0 mL) anddi-tert-butyl dicarbonate (0.68 g, 3.1 mmol) were added and stirred atroom temperature overnight. The reaction was concentrated and theresidue purified by flash chromatography (silica gel, 20% ethylacetate/hexanes) to afford the title compound (0.43 g, 75%) as anoff-white solid.

¹H NMR (300 MHz, DMSO-d₆) δ 13.1 (br. s, 1H), 9.05 (br. s, 2H), 7.93 (d,2H), 7.52 (d, 2H), 1.31 (s, 9H), 1.25 (s, 9H).

Example 11 2-methyl-2-propanyl4-[({(2S,4S)-1-[4-(N′,N″-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidamido)benzoyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 6 was treated with the compound prepared in Example 10 to givethe title compound as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 9.52 (s, 1H), 7.92 (d, 2H),7.61 (d, 2H), 7.57 (d, 2H), 7.22 (d, 2H), 5.00 (t, 1H), 3.94-3.90 (m,1H), 3.45 (t, 1H), 3.29-3.21 (m, 5H), 2.88-2.81 (m, 1H), 2.76 (s, 3H),2.68-2.59 (m, 4H), 2.54-2.46 (m, 2H), 2.41-2.34 (m, 1H), 1.59 (s, 9H),1.57 (s, 18H).

Example 124-[({2S,4S)-1-(4-carbamimidamidobenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 11 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, D20, rotamers present) δ 8.01 (d, 1H), 7.90 (d, 1H),7.68 (d, 1H), 7.62 (d, 1H), 7.49 (d, 1H), 7.42 (d, 1H), 7.23-7.20 (m,2H), 4.84-4.74 (m, 1H), 4.44-4.39 (m, 0.5H), 4.18-4.09 (m, 1H),3.98-3.95 (m, 1H), 3.87-3.82 (m, 0.5H), 3.66-3.11 (m, 8H), 3.05 (s,1.5H), 3.01 (m, 1.5H), 3.00-2.95 (m, 1H), 2.38-2.29 (m, 1H).

ESI MS m/z 558 (M+H)⁺

Example 13 2-methyl-2-propanyl4-[({(2S,4S)-1-({cis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 6 was treated withcis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexanecarboxylicacid to give the title compound as a white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.89 (d, 2H), 7.66 (d, 2H),4.47 (t, 1H), 4.12-4.04 (m, 1H), 3.48 (t, 2H), 3.33 (t, 4H), 3.24-3.22(m, 2H), 2.83 (s, 3H), 2.79 (s, 1H), 2.70-2.59 (m, 6H), 2.01 (s, 1H),1.92-1.80 (m, 6H), 1.58 (s, 9H), 1.43 (s, 9H), 1.23-1.09 (m, 2H).

Example 144-[({(2S,4S)-1-{[cis-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 13 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.97 (d, 2H), 7.71 (d, 2H),4.55 (t, 1H), 4.17 (t, 1H), 4.19-4.16 (m, 1H), 3.68-3.65 (m, 1H),3.54-3.53 (m, 6H), 3.07-3.05 (m, 4H), 3.01-2.98 (m, 4H), 2.92-2.89 (m,2H), 2.05-2.02 (m, 2H), 1.37-1.29 (m, 7H).

ESI MS m/z 536 (M+H)⁺

Example 15 2-methyl-2-propanyl4-[({(2S,4S)-1-({trans-4-[(1S)-1-({[(2-methyl-2-propanyl)oxy]carbonyl}amino)ethyl]cyclohexyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 6 was treated withtrans-4-[(1S)-1-({[(2-methyl-2-propanyl)oxy]carbonyl}amino)ethyl]cyclohexanecarboxylicacid to give the title compound as a light brown solid.

¹H NMR (400 MHz, CDCl₃) δ 9.46 (s, 1H), 7.90 (d, 2H), 7.52 (d, 2H), 4.77(t, 1H), 4.34 (d, 11H), 3.86-3.82 (m, 1H), 3.53-3.51 (m, 11H), 3.37 (t,1H), 3.24-3.22 (m, 4H), 2.95-2.88 (m, 1H), 2.78 (s, 3H), 2.74-2.70 (m,2H), 2.69-2.56 (m, 4H), 2.35-2.34 (m, 1H), 2.26-2.19 (m, 1H), 1.92-1.81(m, 4H), 1.62 (s, 9H), 1.58-1.49 (m, 2H), 1.48 (s, 9H), 1.31-1.24 (m,1H), 1.10 (d, 3H), 1.06-1.04 (m, 1H).

Example 164-[({(2S,4S)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 15 was treated following the proceduredescribed in Example 9 to give the title compound as a brown solid.

¹H NMR (400 MHz, CD₃OD) δ 7.98 (d, 2H), 7.70 (d, 2H), 4.54 (t, 1H),4.25-4.20 (m, 1H), 3.76 (t, 1H), 3.59-3.55 (m, 1H), 3.43-3.34 (m, 4H),3.29-3.08 (m, 6H), 2.91 (s, 3H), 2.70-2.65 (m, 1H), 2.60-2.54 (m, 1H),2.16-2.11 (m, 1H), 2.09-2.03 (m, 1H), 1.91-1.84 (m, 4H), 1.58-1.42 (m,3H), 1.27 (d, 3H).

ESI MS m/z 550 (M+H)⁺

Example 17 methyl4-(N-{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)benzoate

To a solution of methyl 4-(N-carbamimidoyl)benzoate (2.18 g, 9.18 mmol)and triethylamine (1.02 mL, 7.32 mmol) in anhydrous methanol (100 mL)was added di-tert-butyl dicarbonate (3.0 g, 13.8 mmol). The mixture washeated at 40° C. under nitrogen for 5 h. The reaction mixture was cooledand then concentrated under reduced pressure. The resulting residue wasdiluted with ethyl acetate (100 mL) and washed with aqueous sodiumbicarbonate solution. The aqueous layer was then extracted withdichloromethane (2×30 mL). The combined organic extracts were dried andconcentrated. Purification by flash chromatography (silica gel, 80 g,0-30% ethyl acetate/hexanes) afforded the title compound (1.98 g, 77%)as a white solid.

¹H NMR (500 MHz, CDCl₃, Amidine NH protons were not observed.) δ 8.09(d, 2H), 7.91 (d, 2H), 3.94 (s, 3H), 1.55 (s, 9H).

Example 184-(N-{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)benzoic acid

To a solution of the compound prepared in Example 17 (0.20 g, 0.72 mmol)in methanol (10 mL) was added 1 M aqueous sodium hydroxide (5 mL). Thereaction was stirred at room temperature for 1 h. The mixture wasconcentrated and the resulting aqueous residue was diluted with ethylacetate. The aqueous layer was acidified with 1 M hydrochloric acid topH 4-5 and extracted with ethyl acetate (2×20 mL). The combined organicextracts were dried and concentrated to give the title compound (0.208g, >99%) as a white solid.

¹H NMR (500 MHz, DMSO-d₆) δ 8.98 (app. s, 3H), 8.00 (app. s, 4H), 1.44(s, 9H).

ESI MS m/z 263 (M+H)⁺

Example 19 2-methyl-2-propanyl4-[({(2S,4S)-1-[4-(N-{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)benzoyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 6 was treated with the compound prepared in Example 18 to givethe title compound as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 9.47 (s, 1H), 7.93-7.89 (m,4H), 7.61-7.56 (m, 4H), 4.97 (t, 1H), 3.69-3.64 (m, 1H), 3.41 (t, 1H),3.28-3.19 (m, 4H), 2.87-2.80 (m, 1H), 2.77 (s, 3H), 2.65-2.60 (m, 3H),2.48-2.34 (m, 3H), 1.57 (s, 9H), 1.54 (s, 9H).

Example 204-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 19 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, D₂O, rotamers present) δ 9.47 (s, 1H), 8.01-7.61 (m,7H), 7.17 (d, 1H), 4.84-4.76 (m, 1H), 4.09-3.81 (m, 2H), 3.62-3.44 (m,6H), 3.43-3.25 (m, 2H), 3.04-3.00 (m, 4H), 2.36-2.29 (m, 1H).

ESI MS m/z 543 (M+H)⁺

Example 21 methyl trans-4-carbamoylcyclohexanecarboxylate

To a cooled (−10° C.) solution oftrans-4-(methoxycarbonyl)cyclohexane-1-carboxylic acid (1.01 g, 5.43mmol) in tetrahydrofuran (25 mL) was sequentially added triethylamine(1.50 mL, 10.9 mmol) and ethyl chloroformate (0.60 mL, 6.2 mmol) under anitrogen atmosphere and the reaction mixture stirred at room temperaturefor 3 h. The reaction was then cooled to −10° C., ammonium hydroxide(5.0 mL, 33 mmol) was added and the reaction mixture warmed to roomtemperature and stirred overnight whereupon the mixture was diluted withwater and extracted with ethyl acetate (3×50 mL). The combined organicextracts were dried and concentrated to give the title compound (0.84 g,84%) as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 7.17 (s, 1H), 6.66 (s, 1H), 3.58 (s, 3H),2.29-2.20 (m, 1H), 2.07-1.98 (m, 1H), 1.94-1.86 (m, 2H), 1.81-1.71 (m,2H), 1.39-1.23 (m, 4H).

Example 22 methyl trans-4-cyanocyclohexanecarboxylate

To a solution of the compound prepared in Example 21 (0.84 g, 4.6 mmol)in pyridine (20 mL) was added imidazole (1.0 g, 4.6 mmol) andphosphorusoxychloride (1.0 mL) in one portion at 0° C. under a nitrogenatmosphere. The reaction mixture was stirred at room temperature for 4 hwhereupon the reaction was quenched with water and extracted with ethylacetate. The organic extract was washed with 2 M hydrochloric acid,dried and concentrated to give the title compound (0.76 g, 99%) as awhite solid.

¹H NMR (300 MHz, DMSO-d₆) δ 3.58 (s, 3H), 2.71 (t, 1H), 2.38 (t, 1H),2.05-1.80 (m, 4H), 1.63-1.29 (m, 4H).

Example 23 trans-4-cyanocyclohexanecarboxylic acid

To a solution of the compound prepared in Example 22 (0.200 g, 1.19mmol) in tetrahydrofuran (5.0 mL), methanol (1.0 mL) and water (5.0 mL)at 0° C. was added lithium hydroxide (0.073 g, 1.79 mmol). The reactionwas warmed to room temperature and stirred overnight whereupon themixture was carefully acidified to pH 5.0 with 2 N hydrochloric acid andextracted with ethyl acetate (2×300 mL). The combined organic extractswere dried and concentrated to give the title compound (0.15 g, 82%) asa white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 12.14 (br. s, 1H), 2.72-2.65 (m, 1H),2.29-2.22 (m, 1H), 2.0-1.95 (m, 2H), 1.90-1.82 (m, 2H), 1.57-1.47 (m,2H), 1.41-1.34 (m, 2H).

Example 24 2-methyl-2-propanyl4-[({(2S,4S-1-[(trans-4-cyanocyclohexyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 23 was treated with the compound prepared in Example 6 to givethe title compound as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 10.20 (s, 1H), 7.83 (d, 2H),7.67 (d, 2H), 4.34 (t, 1H), 4.03 (t, 1H), 3.10 (m, 5H), 2.86 (s, 3H),2.73-2.56 (m, 4H), 2.05-2.01 (m, 2H), 1.90-1.61 (m, 4H), 1.53 (s, 11H),1.41-1.15 (m, 4H), 1.01-0.96 (m, 1H).

Example 25 ethyl4-[({(2S,4S)-1-[(trans-4-carbamimidoylcyclohexyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoatehydrochloride

Anhydrous HCl gas was bubbled into a solution of the compound preparedin Example 24 (0.11 g, 0.18 mmol) in ethanol (15 mL) at 0° C. for 30 minand the reaction mixture was stirred overnight. 7 M ammonia in methanolwas added to the reaction mixture. After stirring for 5 h, the excessammonia was evaporated to afford the title compound (0.070 g, 65%) whichwas used without further purification.

ESI MS m/z 577 (M+H)⁺

Example 264-[({(2S,4S)-1-[(trans-4-carbamimidoylcyclohexyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

To a solution of the compound prepared in Example 25 (0.070 g, 0.12mmol) in water (0.5 mL) was added 2 M hydrochloric acid (1.0 mL) at 0°C. and the reaction was warmed to room temperature and stirred for 4 h.Solvent was evaporated and the crude compound was purified by Prep-HPLC(0.1% TFA containing CH₃CN—H₂O gradient) to provide the title compound(0.014 g, 21%) as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) 7.98 (d, 2H), 7.70 (d, 2H),4.54 (t, 1H), 4.22-4.18 (m, 1H), 3.70 (t, 1H), 3.50-3.35 (m, 5H),3.14-2.99 (m, 4H), 2.9 (s, 3H), 2.73-2.65 (m, 2H), 2.49-2.33 (m, 1H),2.10-1.96 (m, 5H), 1.67-1.54 (m, 4H).

ESI MS m/z 549 (M+H)⁺

Example 27 benzyl4-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-1-piperazinecarboxylate

Following the procedure described in Example 7, the reaction ofN,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}-1H-pyrazole-1-carboximidamide(0.25 g, 1.1 mmol), triethylamine (0.5 mL, 3.4 mmol) and benzylpiperazine-1-carboxylate (0.422 g, 1.36 mmol) in methanol (10 mL) gavethe title compound (0.256 g, 50%) as a colorless liquid.

¹H NMR (400 MHz, DMSO-d₆) δ 9.63 (br. s, 1H), 7.52-7.34 (m, 5H), 5.10(s, 2H), 3.42-3.40 (m, 8H), 1.36 (s, 18H).

Example 28bis(2-methyl-2-propanyl)(1-piperazinylmethylylidene)biscarbamate

To a solution of the compound prepared in Example 27 (0.045 g, 0.056mmol) in ethanol (10 mL) was added Pd/C (20% by wt, 0.010 g). Thereaction was stirred under an atmosphere of hydrogen (40 psi) at roomtemperature for 2 h. The reaction mixture was filtered throughdiatomaceous earth and the filtrate concentrated to afford the titlecompound (0.03 g, 69%) as a pale green solid.

¹H NMR (400 MHz, DMSO-d₆) δ 9.53 (br. s, 1H), 5.09 (s, 1H), 3.42-3.41(m, 4H), 2.66-2.64 (m, 4H), 1.43 (s, 18H).

Example 29 2-methyl-2-propanyl4-[({(2S,4S)-1-{[4-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-1-piperazinyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

To a solution of the compound prepared in Example 28 (0.05 g, 0.15 mmol)in THF (10 mL), triphosgene (0.054 g, 0.182 mmol) andN,N-diisopropylethylamine (0.08 mL, 0.456 mmol) were added at 0° C. andthe reaction mixture warmed to room temperature. After stirring for 1 h,the reaction mixture was concentrated to dryness and the crude residuewas dissolved in THF. To this mixture, the compound prepared in Example6 (0.065 g, 0.152 mmol) and N,N-diisopropylethylamine (0.08 mL, 0.456mmol) were added at 0° C., and the reaction warmed to room temperature.After stirring for 4 h, the mixture was diluted with ice cold water andextracted with ethyl acetate (2×50 mL). The organic layer was washedwith brine, dried and concentrated. Purification by flash chromatography(silica gel, 40 g, 20-60% ethyl acetate/hexanes) provided the titlecompound (0.045 g, 39%) as a white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.89 (d, 2H), 7.67 (d, 2H),4.65-4.62 (m, 1H), 4.11-4.09 (m, 1H), 3.75-3.71 (m, 2H), 3.52-3.49 (m,8H), 3.23 (t, 4H), 2.86-2.85 (m, 1H), 2.83 (s, 3H), 2.71-2.62 (m, 5H),1.61 (s, 9H), 1.47 (s, 18H).

Example 304-[({(2S,4S)-1-[(4-carbamimidoyl-1-piperazinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 29 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.84 (d, 2H), 7.71 (d, 21H),4.69-4.67 (m, 1H), 3.88-3.84 (m, 1H), 3.64-3.55 (m, 5H), 3.54-3.52 (m,2H), 3.50-3.49 (m, 2H), 3.48-3.38 (m, 5H), 3.03-3.02 (m, 2H), 2.99-2.89(s, 3H), 2.72 (m, 2H), 1.37-1.12 (m, 2H).

ESI MS m/z 551 (M+H)⁺

Example 31 2-methyl-2-propanyl4-[({(2S,4S)-1-({trans-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

Following the procedure described in Example 8, the compound prepared inExample 6 was treated withtrans-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexanecarboxylicacid to give the title compound as a white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.89 (d, 2H), 7.66 (d, 2H),4.47 (t, 1H), 4.12-4.04 (m, 1H), 3.48 (t, 2H), 3.33 (t, 4H), 3.24-3.22(m, 2H), 2.83 (s, 3H), 2.79 (s, 1H), 2.70-2.59 (m, 6H), 2.01 (s, 1H),1.92-1.80 (m, 6H), 1.58 (s, 9H), 1.43 (s, 9H), 1.23-1.09 (m, 2H).

Example 324-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 31 was treated following the proceduredescribed in Example 9 to give the title compound as a white solid.

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.59 (t, 1H), 4.37-4.26 (m, 1H), 3.99-3.84 (m, 2H), 3.54 (br. s, 4H),3.43 (br. s, 4H), 2.96 (s, 3H), 2.88-2.77 (m, 3H), 2.60 (tt, 1H),2.37-2.22 (m, 1H), 2.03-1.96 (m, 1H), 1.96-1.83 (m, 3H), 1.69-1.56 (m,1H), 1.56-1.41 (m, 21H), 1.21-1.10 (m, 2H).

ESI MS m/z 536 (M+H)⁺

Example 334-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6, 31 and 32 to give the title compoundhaving the following physical properties. (Note: in the stepcorresponding to Example 3 in the operation, morpholine was used inplace of 1-(methylsulfonyl)piperazine)

¹H NMR (250 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.60 (t, 1H), 4.43-4.28 (m, 1H), 4.15-3.79 (m, 6H), 3.44 (br. s, 4H),2.92-2.76 (m, 3H), 2.75-2.50 (m, 1H), 2.41-2.24 (m, 1H), 2.07-1.80 (m,4H), 1.80-1.35 (m, 3H), 1.27-1.02 (m, 2H).

ESI MS m/z 459 (M+H)⁺

Example 344-({[(3′S,5′S)-1′-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-1,3′-bipyrrolidin-5′-yl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6, 31 and 32 to give the title compoundhaving the following physical properties. (Note: in the stepcorresponding to Example 3 in the operation, pyrrolidine was used inplace of 1-(methylsulfonyl)piperazine).

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.72 (d, 2H),4.62 (dd, 1H), 4.26 (dd, 1H), 4.06 (quint., 1H), 3.98 (dd, 1H), 3.73(br. s, 2H), 3.28 (br. s, 2H), 2.87-2.77 (m, 3H), 2.59 (tt, 1H),2.38-2.31 (m, 1H), 2.14 (br. s, 4H), 2.01-1.83 (m, 4H), 1.69-1.58 (m,1H), 1.53-1.41 (m, 2H), 1.21-1.09 (m, 2H).

ESI MS m/z 443 (M+H)⁺

Example 354-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(1-piperidinyl)-2-pyrrolidinyl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6, 31 and 32 to give the title compoundhaving the following physical properties. (Note: in the stepcorresponding to Example 3 in the operation, piperidine was used inplace of 1-(methylsulfonyl)piperazine).

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.57 (t, 1H), 4.37 (dd, 1H), 3.99 (quint., 1H), 3.87 (t, 1H), 3.65 (br.s, 2H), 3.05 (br. s, 2H), 2.87 (ddd, 1H), 2.80 (d, 2H), 2.60 (tt, 1H),2.29-2.20 (m, 1H), 2.07-1.71 (m, 9H), 1.69-1.57 (m, 1H), 1.57-1.39 (m,3H), 1.22-1.09 (m, 2H).

ESI MS m/z 457 (M+H)⁺

Example 364-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(3-oxo-1-piperazinyl)-2-pyrrolidinyl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6, 31 and 32 to give the title compoundhaving the following physical properties. (Note: in the stepcorresponding to Example 3, 2-oxopiperazine was used in place of1-(methylsulfonyl)piperazine).

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.56 (t, 1H), 4.24 (dd, 1H), 3.81 (dd, 1H), 3.75-3.60 (m, 3H), 3.50 (dd,2H), 3.39-3.31 (m, 1H), 3.27-3.19 (m, 1H), 2.80 (d, 2H), 2.80-2.71 (m,1H), 2.60 (tt, 1H), 2.20 (dt, 1H), 2.03-1.95 (m, 1H), 1.95-1.83 (m, 3H),1.69-1.58 (m, 1H), 1.54-1.40 (m, 2H), 1.21-1.09 (m, 2H).

ESI MS m/z 472 (M+H)⁺

Example 374-({[(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoicacid trifluoroacetate

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6, 8 and 9 to give the title compoundhaving the following physical properties. (Note: in the stepcorresponding to Example 3, morpholine was used in place of1-(methylsulfonyl)piperazine. In the step corresponding to Example 8,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid was used inplace of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 9.53 (s, 1H), 8.15 (d, 1H),7.98 (d, 2H), 7.70 (d, 2H), 7.68 (dd, 1H), 7.59 (d, 1H), 7.18 (d, 1H),7.05 (d, 1H), 4.70 (t, 1H), 4.41 (dd, 1H), 4.17-3.84 (m, 6H), 3.45 (br.s, 4H), 2.90-2.81 (m, 1H), 2.46-2.36 (m, 1H).

ESI MS m/z 552 (M+H)⁺

Example 38 methyl4-({[(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinyl]carbonyl}amino)benzoate

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, methyl 4-aminobenzoate was usedin place of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.38 (s, 1H), 9.85 (s,1H), 8.36 (d, 1H), 7.91 (d, 2H), 7.76 (dd, 1H), 7.74-7.67 (m, 3H), 7.28(d, 1H), 6.85 (d, 1H), 4.44 (dd, 1H), 4.28 (dd, 11H), 3.82 (s, 3H),3.65-3.55 (m, 4H), 3.41 (t, 1H), 2.93-2.83 (m, 1H), 2.52-2.40 (m, 51H),1.70 (q, 1H).

ESI MS m/z 566 (M+H)⁺

Example 39(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-4-[4-(phenylsulfonyl)-1-piperazinyl]-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Example 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, 1-benzenesulfonyl-piperazine was used in place of1-(methylsulfonyl)piperazine. In the step corresponding to Example 5,aniline was used in place of tert-butyl 4-aminobenzoate. In the stepcorresponding to Example 8,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid was used inplace of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.02 (s, 1H), 9.85 (s,1H), 8.34 (d, 1H), 7.78-7.69 (m, 5H), 7.67 (t, 2H), 7.52 (d, 2H),7.30-7.22 (m, 3H), 7.01 (t, 1H), 6.83 (d, 1H), 4.40 (t, 1H), 4.26 (dd,1H), 3.31 (dd, 1H), 2.94-2.85 (m, 5H), 2.62-2.49 (m, 4H), 2.47-2.38 (m,1H), 1.60 (dd, 1H).

ESI MS m/z 647 (M+H)⁺

Example 40(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-[4-(methylsulfonyl)phenyl]-4-(4-morpholinyl)-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 4-(methanesulfonyl)aniline wasused in place of tert-butyl 4-aminobenzoate. In the step correspondingto Example 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acidwas used in place of 1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.49 (s, 1H), 9.85 (s,1H), 8.36 (d, 1H), 7.85 (d, 2H), 7.81 (d, 2H), 7.76 (dd, 1H), 7.72 (d,1H), 7.28 (d, 1H), 6.85 (d, 1H), 4.44 (t, 1H), 4.29 (dd, 1H), 3.60 (br.s, 4H), 3.41 (t, 1H), 3.15 (s, 3H), 2.94-2.84 (m, 1H), 2.46 (app. br. s,5H), 1.70 (dd, 11H).

ESI MS m/z 586 (M+H)⁺

Example 41(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-N-(3-pyridinyl)-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 3-aminopyridine was used inplace of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.26 (s, 1H), 9.86 (s,1H), 8.72 (d, 1H), 8.36 (d, 1H), 8.25 (d, 1H), 8.01 (d, 1H), 7.76 (dd,1H), 7.72 (d, 1H), 7.33 (dd, 1H), 7.28 (d, 1H), 6.86 (d, 1H), 4.43 (t,1H), 4.28 (dd, 1H), 3.60 (br. s, 4H), 3.41 (t, 1H), 2.93-2.83 (m, 1H),2.53-2.30 (m, 5H), 1.71 (dd, 1H).

ESI MS m/z 509 (M+H)⁺

Example 42(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-(4-methoxyphenyl)-4-(4-morpholinyl)-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 4-methoxyaniline was used inplace of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) & 9.87 (s, 1H), 9.86 (s,1H), 8.36 (d, 1H), 7.76 (dd, 1H), 7.71 (d, 1H), 7.47 (d, 2H), 7.28 (d,1H), 6.86 (d, 2H), 6.85 (d, 1H), 4.38 (t, 1H), 4.27 (dd, 1H), 3.71 (s,3H), 3.63-3.57 (m, 4H), 3.38 (obs. t, 1H), 2.88-2.79 (m, 1H), 2.53-2.39(m, 5H), 1.68 (dd, 1H).

ESI MS m/z 538 (M+H)⁺

Example 43(2S,4S)—N-(1H-benzotriazol-6-yl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamidetrifluoroacetate

The compound prepared in Example 2 was treated following the proceduresdescribed in Example 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 5-aminobenzotriazole was used inplace of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 9.53 (s, 1H), 8.35 (d, 1H),8.16 (d, 1H), 7.84 (d, 1H), 7.68 (dd, 1H), 7.59 (d, 1H), 7.46 (dd, 1H),7.19 (d, 1H), 7.07 (d, 1H), 4.74 (t, 1H), 4.42 (dd, 1H), 4.12 (dd, 1H),4.09-4.02 (m, 1H), 3.98 (br. s, 4H), 3.47 (br. s, 4H), 2.91-2.83 (m,1H), 2.49-2.41 (m, 1H).

ESI MS m/z 549 (M+H)⁺

Example 44(2S,4S)—N-(3-chlorophenyl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 3-chloroaniline was used inplace of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.21 (s, 1H), 9.86 (s,1H), 8.35 (d, 1H), 7.78 (app. s, 1H), 7.75 (dd, 1H), 7.71 (d, 1H), 7.41(d, 1H), 7.33 (t, 1H), 7.28 (d, 1H), 7.10 (d, 18H), 6.86 (d, 1H), 4.39(t, 1H), 4.27 (dd, 1H), 3.58 (br. s, 4H), 3.39 (t, 1H), 2.94-2.82 (m,1H), 2.51-2.38 (m, 5H), 1.68 (dd, 1H).

ESI MS m/z 542/544 (M+H)⁺

Example 45(2S,4S)—N-(3-chloro-4-fluorophenyl)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 3-chloro-4-fluoroaniline wasused in place of tert-butyl 4-aminobenzoate. In the step correspondingto Example 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acidwas used in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.27 (s, 1H), 9.86 (s,1H), 8.32 (d, 1H), 7.90 (dd, 1H), 7.75 (dd, 1H), 7.71 (d, 1H), 7.48-7.40(m, 1H), 7.36 (t, 1H), 7.27 (d, 1H), 6.85 (d, 1H), 4.38 (t, 1H), 4.27(dd, 1H), 3.58 (br. s, 4H), 3.41 (t, 1H), 2.93-2.79 (m, 1H), 2.52-2.39(m, 5H), 1.68 (dd, 1H).

ESI MS (ES⁺) m/z 560/562 (M+H)⁺

Example 46(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-(4-morpholinyl)-N-(2-oxo-2,3-dihydro-1H-benzimidazol-5-yl-2-pyrrolidinecarboxamidetrifluoroacetate

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, morpholine was used in place of 1-(methylsulfonyl)piperazine.In the step corresponding to Example 5, 5-aminobenzimidazolone was usedin place of tert-butyl 4-aminobenzoate. In the step corresponding toExample 8, (2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid wasused in place of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.68 (s, 1H), 10.52 (s,1H), 10.37-9.93 (br. s, 1H), 9.87 (s, 1H), 8.29 (app. br. s, 1H), 7.78(dd, 1H), 7.75 (d, 1H), 7.39 (s, 1H), 7.26 (d, 1H), 6.99 (d, 1H),6.94-6.73 (m, 2H), 4.48 (app. br. s, 1H), 4.32-3.48 (m, 71H), 3.26-2.15(m, 5H), 2.18-1.94 (m, 1H).

ESI MS m/z 564 (M+H)⁺

Example 47(2S,4S)-1-[(3-chloro-1H-indol-5-yl)carbonyl]-4-[4-(cyclopropylsulfonyl)-1-piperazinyl]-N-phenyl-2-pyrrolidinecarboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 6 and 8 to give the title compound havingthe following physical properties. (Note: in the step corresponding toExample 3, 1-(cyclopropanesulfonyl)piperazine was used in place of1-(methylsulfonyl)piperazine. In the step corresponding to Example 5,aniline was used in place of tert-butyl 4-aminobenzoate. In the stepcorresponding to Example 8, 3-chloroindole-5-carboxylic acid was used inplace of1-(N,N′-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidoyl)-4-piperidinecarboxylicacid).

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 9.43 (s, 1H), 8.53 (s, 1H),7.86 (s, 1H), 7.57 (d, 2H), 7.43 (d, 1H), 7.40 (d, 1H), 7.32-7.25 (m,3H), 7.08 (t, 1H), 5.105.00 (m, 1H), 3.93-3.84 (m, 1H), 3.59-3.50 (m,1H), 3.31 (br. s, 4H), 2.88-2.78 (m, 1H), 2.73-2.57 (m, 3H), 2.53-2.44(m, 2H), 2.43-2.35 (m, 1H), 2.28-2.20 (m, 1H), 1.19-1.13 (m, 2H),1.02-0.95 (m, 2H).

ESI MS m/z 556 (M+H)⁺

Example 48 methyl4-({[(2S,4S)-4-amino-1-({trans-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-2-pyrrolidinyl]carbonyl}aminobenzoate

Methyl-4-[(2S,4S)-1-{([4-({[(tert-butoxy)carbonyl]amino}methyl)cyclohexyl]carbonyl}-4-{[(9H-fluoren-9-ylmethoxy)carbonyl]amino}pyrrolidine-2-amido]benzoatewas synthesized by following the procedures described in Examples 5, 9and 8 starting from(2S,4S)-1-tert-butoxycarbonyl-4-(9-fluorenylmethoxycarbonyl)amino-pyrrolidine-2-carboxylicacid. (Note: in the step corresponding to Example 5, methyl4-aminobenzoate was used in place of tert-butyl 4-aminobenzoate. In thestep corresponding to Example 8,trans-4-(tert-butoxycarbonylamino)cyclohexanecarboxylic acid was used inplace of the compound prepared in Example 7) The crude material (11.61g) was suspended in anhydrous tetrahydrofuran (120 mL) and piperidine(6.6 mL) added dropwise. The reaction was stirred for 2 hours thenfurther piperidine (6.6 mL) was added and the reaction stirredovernight. The solvent was removed in vacuo and the residue purified bycolumn chromatography (silica gel, 25-100% ethyl acetate/heptanes, 5-10%methanol/ethyl acetate then 0-20% methanol/dichloromethane) to give thetitle compound (3.04 g) as pale yellow foam.

ESI MS m/z 503 (M+H)⁺

Example 49 methyl4-({[(2S,4S)-4-(4-benzyl-1-piperazinyl)-({cis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-2-pyrrolidinyl]carbonyl}amino)benzoate

To a solution of the compound prepared in Example 48 (1.05 g) in2-propanol (56 mL) was added N-benzyl-N,N-bis(2-chloroethyl)aminehydrochloride (0.59 g) and sodium bicarbonate (3.6 g) and the reactionrefluxed for 16 h. The reaction was reduced in vacuo, the residuediluted with water and extracted with ethyl acetate. The combined ethylacetate fractions were washed with brine, dried and concentrated. Theresidue was purified by column chromatography (silica gel, 0-20%methanol/dichloromethane) to give the title compound (0.90 g) as acolourless solid.

ESI MS m/z 662 (M+H)⁺

Example 50 methyl4-({[(2S,4S)-1-({cis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-4-(1-piperazinyl)-2-pyrrolidinyl]carbonyl}amino)benzoate

To a flask under nitrogen was added the compound prepared in Example 49(0.90 g), ammonium formate (2.57 g) and palladium on carbon (1.61 g).Methanol was added (45 mL) under nitrogen and the reaction heated atreflux for 2.5 h. The reaction mixture was cooled and filtered throughCelite®, washing the filter cake with methanol. The solvent was removedin vacuo, diluted with water and extracted with ethyl acetate. Thecombined ethyl acetate fractions were washed with brine, dried andconcentrated to give the title compound (0.57 g) as a colourless solid.

ESI MS m/z 572 (M+H)⁺

Example 51 methyl4-[(3S,5S)-5-{[4-(methoxycarbonyl)phenyl]carbamoyl}-1-({cis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexyl}carbonyl)-3-pyrrolidinyl]-1-piperazinecarboxylate

To a stirred solution of the compound prepared in Example 50 (150 mg) indichloromethane (3 mL) was added triethylamine (0.3 mL) and methylchloroformate (22 μL). The mixture was stirred at room temperatureovernight. Further methyl chloroformate (11 μL) was added and thereaction stirred at room temperature for an additional 4 days. Thereaction mixture was partitioned between dichloromethane (20 mL) andbrine (20 mL), the organic layer separated, dried and concentrated. Theresidue was purified by column chromatography (silica gel, ethylacetate) to give the title compound (119 mg) as a white solid.

ESI MS m/z 630 (M+H)⁺

Example 524-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(methoxycarbonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

To a solution of the compound prepared in Example 51 (119 mg) inmethanol (1 mL) and THF (1 mL) was added 1 N sodium hydroxide (2 mL) andthe mixture stirred at room temperature for 2 h. The organic solventswere removed in vacuo, the residual aqueous phase neutralised byaddition of 1 N hydrochloric acid (2 mL) and extracted intodichloromethane (3×10 mL). The combined organic phases were washed withbrine, dried and concentrated. The residue (99 mg) was redissolved indichloromethane (5 mL) and treated with trifluoroacetic acid (2 mL). Thereaction mixture was stirred at room temperature for 1 hour thenconcentrated. Purification by high performance liquid chromatography ([5to 100% mobile phase B (0.1% trifluoroacetic acid in acetonitrile) inmobile phase A (0.1% trifluoroacetic acid in water] gave the titlecompound (90 mg, 64%) as a white solid.

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.59 (t, 1H), 4.34 (dd, 1H), 4.04-3.96 (m, 1H), 3.93 (dd, 1H), 3.78 (br.s, 4H), 3.75 (s, 3H), 3.48-3.33 (m, 4H), 2.89-2.81 (m, 1H), 2.80 (d,2H), 2.60 (tt, 1H), 2.32 (dt, 1H), 2.03-1.95 (m, 11H), 1.95-1.83 (m,3H), 1.69-1.55 (m, 11H), 1.55-1.41 (m, 2H), 1.23-1.09 (m, 2H).

ESI MS m/z 516 (M+H)⁺

Example 534-({[(2S,4S)-4-(4-acetyl-1-piperazinyl)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-2-pyrrolidinyl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

The compound prepared in Example 50 was treated following the proceduresdescribed in Example 51 and 52 to give the title compound having thefollowing physical properties. (Note: in the step corresponding toExample 51, acetic anhydride was used in place of methyl chloroformate.)

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.59 (t, 1H), 4.35-4.28 (m, 1H), 3.95-3.76 (m, 6H), 3.39 (br. s, 2H),3.35 (br. s, 2H), 2.88-2.77 (m, 3H), 2.60 (tt, 1H), 2.34-2.25 (m, 1H),2.16 (s, 3H), 2.03-1.96 (m, 1H), 1.96-1.84 (m, 3H), 1.70-1.58 (m, 1H),1.53-1.42 (m, 2H), 1.21-1.09 (m, 2H).

ESI MS m/z 500 (M+H)⁺

Example 544-({[(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-(4-carbamoyl-1-piperazinyl)-2-pyrrolidinyl]carbonyl}amino)benzoicacid bis(trifluoroacetate)

To a solution of the compound prepared in Example 50 (150 mg) in aceticacid (2 mL) and water (2 mL) was added potassium isocyanate (105 mg) andthe mixture stirred at room temperature for 4 days. The reaction mixturewas then treated with saturated aqueous sodium bicarbonate (10 mL) andextracted into dichloromethane (3×10 mL). The combined organic phaseswere washed with brine, dried and concentrated. Purification by flashchromatography (silica gel, 5-10% methanol/dichloromethane) gave methyl4-[(2S,4S)-1-{[4-({[(tert-butoxy)carbonyl]amino}methyl)cyclohexyl]carbonyl}-4-(4-carbamoylpiperazin-1-yl)pyrrolidine-2-amido]benzoate(54 mg) as a colourless oil.

The compound thus obtained was treated following the procedure describedin Example 52 to give the title compound (51 mg, 29%) as a white solid.

¹H NMR (500 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.60 (t, 1H), 4.36 (dd, 1H), 4.08-3.99 (m, 1H), 3.95 (dd, 1H), 3.74 (br.s, 4H), 3.43 (br. s, 4H), 2.91-2.83 (m, 1H), 2.80 (d, 2H), 2.60 (tt,1H), 2.34 (dt, 1H), 2.03-1.95 (m, 1H), 1.95-1.83 (m, 3H), 1.69-1.56 (m,1H), 1.55-1.41 (m, 2H), 1.22-1.10 (m, 2H).

ESI MS m/z 501 (M+H)⁺

Example 554-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(cyclopropylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 50 was treated following the proceduresdescribed in Example 51 and 52 to give the title compound having thefollowing physical properties. (Note: in the step corresponding toExample 51, cyclopropanesulfonyl chloride was used in place of methylchloroformate.)

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 8.03-7.91 (m, 2H), 7.77-7.63(m, 2H), 4.55 (t, 1H), 4.26 (t, 1H), 3.96-3.60 (m, 2H), 3.60-3.40 (m,4H), 3.40-3.09 (m, 4H), 2.88-2.45 (m, 5H), 2.43-2.08 (m, 1H), 2.06-1.35(m, 7H), 1.27-0.96 (m, 6H).

FAB MS m/z 562 (M+H)⁺

Example 55-24-[({(2S,4S)-1-{[trans-4-(aminomethyl)cyclohexyl]carbonyl}-4-[4-(ethylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 50 was treated following the proceduresdescribed in Example 51 and 52 to give the title compound having thefollowing physical properties. (Note: in the step corresponding toExample 51, ethanesulfonyl chloride was used in place of methylchloroformate.)

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 8.06-7.91 (m, 2H), 7.88-7.62(m, 2H), 4.55 (t, 1H), 4.35-4.17 (m, 1H), 3.88-3.61 (m, 2H), 3.61-3.41(m, 4H), 3.41-3.03 (m, 6H), 2.89-2.49 (m, 3H), 2.44-2.08 (m, 1H),2.06-1.25 (m, 11H), 1.25-0.98 (m, 2H).

FAB MS m/z 550 (M+H)⁺

Example 56(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-4-(4-sulfamoyl-1-piperazinyl)-2-pyrrolidinecarboxamidetrifluoroacetate

The compound prepared in Example 2 was treated following the proceduresdescribed in Example 3, 4, 5, 9, 51, 6, 8 and 9 to give the titlecompound having the following physical properties. (Note: in the stepscorresponding to Examples 3, 5, 8 and 51,1-(tert-butoxycarbonyl)piperazine, aniline,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid and sulfamidein 1,4-dioxane were used respectively)

¹H NMR (500 MHz, DMSO-d₆ rotamers present) δ 10.06 (br. s, 1H), 9.86 (s,1H), 8.37 (d, 1H), 7.76 (dd, 1H), 7.72 (d, 1H), 7.56 (d, 2H), 7.34-7.25(m, 3H), 7.03 (t, 1H), 6.85 (d, 1H), 6.75 (br. s, 2H), 4.43 (t, 1H),4.32 (dd, 1H), 3.43-3.34 (obs. m, 1H), 2.98 (br. s, 4H), 2.94-2.85 (m,1H), 2.58 (br. s, 4H), 2.50-2.40 (obs. m, 1H), 1.68 (dd, 1H).

ESI MS m/z 586 (M+H)⁺, 557 [(M-N₂)]

Example 57(2S,4S)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[(3S)-3-methyl-4-sulfamoyl-1-piperazinyl]-N-phenyl-2-pyrrolidinecarboxamidetrifluoroacetate

The compound prepared in Example 2 was treated following the proceduresdescribed in Example 3, 4, 5, 9, 51, 6, 8 and 9 to give the titlecompound having the following physical properties. (Note: in the stepcorresponding to the Example 3, 5, 8 and 51,1-(tert-butoxycarbonyl)-2(S)-methylpiperazine, aniline,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid and sulfamidein 1,4-dioxane were used respectively)

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 9.65 (s, 1H), 8.23-8.00(m, 1H), 7.74-7.60 (m, 2H), 7.56 (d, 2H), 7.27 (t, 2H), 7.13-6.90 (m,3H), 4.73-4.42 (m, 1H), 4.21-4.04 (m, 1H), 3.96-3.46 (m, 4H), 3.44-3.32(m, 1H), 3.31-2.97 (m, 3H), 2.97-2.80 (m, 1H), 2.80-2.65 (m, 1H),1.99-1.72 (m, 1H), 1.32-1.18 (m, 3H).

ESI MS m/z 600 (M+H)⁺

Example 58(3R,3′S,5′S)-1′-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-N-phenyl-3-(sulfamoylamino)-1,3′-bipyrrolidine-5′-carboxamide

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 5, 9, 51, 6, 8 and 9 to give the titlecompound having the following physical properties. (Note: in the stepcorresponding to the Example 3, 5, 8 and 51,(3R)-3-(tert-butoxycarbonylamino)pyrrolidine, aniline,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid and sulfamidein 1,4-dioxane were used respectively)

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 10.01 (s, 1H), 9.85 (s,1H), 8.38 (d, 1H), 7.75 (dd, 1H), 7.71 (d, 1H), 7.55 (d, 2H), 7.31-7.25(m, 3H), 7.03 (t, 1H), 6.85 (d, 11H), 6.72 (d, 1H), 6.53 (br. s, 21H),4.42 (t, 1H), 4.20 (dd, 1H), 3.82-3.71 (m, 1H), 3.48-3.39 (obs. m, 1H),2.99-2.89 (m, 1H), 2.88-2.77 (m, 1H), 2.64-2.51 (obs. m, 3H), 2.48-2.36(m, 2H), 2.14-2.00 (m, 1H), 1.74-1.62 (m, 1H).

ESI MS (ES⁺) m/z 608 (M+Na)⁺, 586 (M+H)⁺

Example 59 2-methyl-2-propanyl4-[({(2S,4S)-1-({4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]-1-piperidinyl}carbonyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

To a solution of the compound prepared in Example 6 (0.050 g, 0.11 mmol)in THF (10 mL) were added triphosgene (0.099 g, 0.33 mmol),(boc-4-aminomethyl)-piperidine (0.060 g, 0.28 mmol) andN,N-diisopropylethylamine (0.146 mL, 0.84 mmol) and the reaction stirredat 0° C. for 2 h. The reaction was quenched by adding ice cold water andthe resulting precipitate was collected by filtration, dried and thecrude product purified by flash chromatography to afford the titlecompound (0.052 g, 33%) as a white solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) 89.12 (s, 1H), 7.93 (d, 2H),7.55 (d, 2H), 4.80 (dd, 1H), 4.62-4.57 (m, 1H), 3.83-3.65 (m, 2H),3.57-3.48 (m, 1H), 3.32 (t, 1H), 3.26-3.24 (m, 4H), 3.04-2.97 (m, 3H),2.89-2.81 (m, 2H), 2.78 (s, 3H), 2.71-2.66 (m, 3H), 2.58-2.53 (m, 2H),2.41-2.46 (m, 2H), 1.76-1.62 (m, 4H), 1.58 (s, 9H), 1.43 (s, 9H).

Example 604-[({(2S,4S)-1-{[4-(aminomethyl)-1-piperidinyl]carbonyl}-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 59 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.98 (d, 2H), 7.71 (d, 2H),4.68 (dd, 1H), 3.94-3.81 (m, 1H), 3.74-3.69 (m, 1H), 3.59-3.39 (m, 4H),3.23-3.08 (m, 4H), 3.03-2.94 (m, 1H), 2.91 (s, 3H), 2.85-2.70 (m, 4H),2.09 (q, 1H), 1.89-1.71 (m, 3H), 1.42-1.14 (m, 5H).

ESI MS m/z 537 (M+H)⁺

Example 61 2-methyl1-(2-methyl-2-propanyl)(2S)-4-(4-pyridinyl)-2,5-dihydro-1H-pyrrole-1,2-dicarboxylate

A solution of 2-methyl 1-(2-methyl-2-propanyl)(2S)-4-(trifluoromethylsulfonyloxy)-1H-pyrrole-1,2(2H,5H)-dicarboxylate(11.3 g, 30.1 mmol) and 4-pyridineboronic acid (4.44 g, 36.1 mmol) in1,4-dioxane (120 mL) was purged with argon gas, followed by the additionof tetrakis(triphenylphosphine)palladium (1.04 g, 0.9 mmol) and sodiumcarbonate (2 M in water, 38 mL, 76 mmol). The reaction was heated at105° C. and stirred under argon atmosphere for 1 h whereupon thereaction was cooled to room temperature and concentrated to a volume of50 mL. The solution was diluted with ethyl acetate (200 mL) and filteredthrough diatomaceous earth with a layer of sodium sulfate on the top.The filtrate was concentrated and purified by flash chromatography(silica gel, 400 g, 20-70% ethyl acetate/hexanes) to afford the titlecompound (6.4 g, 70%) as a pale yellow solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 8.63-8.61 (m, 2H), 7.25-7.23(m, 2H), 6.33 (d, 0.4H), 6.28 (d, 0.6H), 5.23 (dt, 0.4H), 5.16 (dt,0.6H), 4.65-4.55 (m, 2H), 3.78 (s, 1.2H), 3.77 (s, 1.8H), 1.53 (s,3.6H), 1.46 (s, 5.4H).

Example 62 methyl(2S,4R)-1-{[(2-methyl-2-propanyl)oxy]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinecarboxylate

A solution of the compound prepared in Example 61 (4.31 g, 14.1 mmol)and acetic acid (0.8 mL) in ethanol (80 mL) was purged with hydrogen.Platinum oxide (0.86 g, 20 wt %) was added and the mixture heated at 50°C. and stirred overnight under an atmosphere of hydrogen. The reactionmixture was then cooled to room temperature and filtered throughdiatomaceous earth with methanol. The filtrate was concentrated and theresidue partitioned between dichloromethane (100 mL) and water (50 mL).The aqueous layer was adjusted to pH 10 by addition of 2 M sodiumhydroxide and extracted with dichloromethane (4×50 mL). The combinedorganic extracts were concentrated and azeotroped with toluene to givethe crude product as an off-white foam. The crude material was dissolvedin dichloromethane (100 mL) and cooled to 0° C. Triethylamine (3.93 mL,28.2 mmol) was added, followed by methylsulfonyl chloride (1.64 mL, 21.1mmol). The reaction was stirred under nitrogen at 0° C. for 1 h then atroom temperature overnight whereupon the mixture was diluted with ethylacetate (300 mL) and washed with 1 M hydrochloric acid (50 mL), aqueoussodium bicarbonate (50 mL) and brine (50 mL). The organic layer was thendried and concentrated. Purification by flash chromatography (silicagel, 120 g, 0-8% methanol/dichloromethane) afforded the title compound(2.6 g, 47% for two steps) as a pale yellow foam.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 4.26 (dd, 0.3H), 4.20 (dd,0.7H), 3.86-3.78 (m, 2.1H), 3.74 (s, 0.9H), 3.72 (s, 2.11H), 3.72-3.69(m, 0.9H), 3.06 (t, 1H), 2.76 (s, 3H), 2.63-2.59 (m, 2H), 2.45-2.42 (m,1H), 1.98-1.91 (m, 1H), 1.79-1.73 (m, 2H), 1.60-1.52 (m, 1H), 1.46 (s,2.7H), 1.40 (s, 6.3H), 1.46-1.25 (m, 3H).

Example 63(2S,4R)-1-{[(2-methyl-2-propanyl)oxy]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinecarboxylicacid

To a solution of the compound prepared in Example 62 (2.87 g, 7.36 mmol)in tetrahydrofuran (20 mL) and water (20 mL) at 0° C. was added lithiumhydroxide (0.35 g, 14.7 mmol). The reaction was warmed to roomtemperature and stirred overnight whereupon it was partitioned betweenmethyl tert-butyl ether and water. The aqueous layer was separated andcarefully acidified to pH 1 with 6 N hydrochloric acid. The aqueoussolution was extracted with ethyl acetate (2×300 mL) and the combinedorganic extracts concentrated to give the title compound (2.35 g, 85%)as a white solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 4.35 (t, 0.7H), 4.27-4.23(m, 0.3H), 3.83-3.73 (m, 3H), 3.10-2.97 (m, 1H), 2.77 (s, 3H), 2.62 (td,2H), 2.54-2.47 (m, 0.3H), 2.36-2.32 (m, 0.7H), 2.13-2.06 (m, 0.7H),1.96-1.89 (m, 1.3H), 1.77-1.73 (m, 1H), 1.50 (s, 6.3H), 1.42 (s, 2.7H),1.45-1.32 (m, 4H).

Example 64 2-methyl-2-propanyl(2S,4R)-2-(5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinecarboxylate

To a solution of the compound prepared in Example 62 (2.35 g, 6.2 mmol)in N,N-dimethylformamide (50 mL) at 0° C. was added methyl4-(2-bromoacetyl)phenylcarbamate (2.04 g, 7.49 mmol) followed by cesiumcarbonate (4.47 g, 13.7 mmol). The reaction was warmed to roomtemperature and stirred for 1.5 h whereupon the mixture was filteredthrough diatomaceous earth and the filter cake washed withdichloromethane. The filtrate was concentrated to afford crude productas a yellow solid. To a suspension of crude material in xylene (45 mL)in a glass pressure bottle was added ammonium acetate (2.88 g, 37.4mmol). The reaction vessel was filled with nitrogen, sealed and heatedat 140° C. for 1 h whereupon the mixture was cooled to room temperature,concentrated and partitioned between ethyl acetate (200 mL) and water(50 mL). The organic layer was washed with aqueous sodium bicarbonatesolution and brine, dried and concentrated. Purification by flashchromatography (silica gel, 120 g, 40-60% ethyl acetate/dichloromethanethen 3-5% methanol/dichloromethane) afforded the title compound (2.42 g,71% for two steps) as a pale yellow solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 11.06 (s, 0.4H), 10.66 (s,0.6H), 7.70-7.67 (m, 2H), 7.40-7.36 (m, 2H), 7.19 (s, 0.6H), 7.13 (s,0.4H), 6.71 (s, 0.4H), 6.66 (s, 0.6H), 4.97-4.91 (m, 1H), 3.83-3.77 (m,3H), 3.77 (s, 3H), 2.96-2.94 (m, 1H), 2.76 (s, 3H), 2.62-2.59 (m, 2H),2.56-2.50 (m, 1H), 2.14-2.11 (m, 0.6H), 2.00-1.92 (m, 1.4H), 1.78-1.75(m, 1H), 1.48 (s, 9H), 1.45-1.38 (m, 4H).

ESI MS m/z 548 (M+H)⁺

Example 65 2-methyl-2-propanyl(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinecarboxylate

To a solution of the compound prepared in Example 64 (2.41 g, 4.4 mmol)in acetonitrile (50 mL) and N,N-dimethylformamide (20 mL) at 0° C. wasadded N-chlorosuccinimide (0.62 g, 4.6 mmol). The reaction was warmed toroom temperature, stirred under nitrogen overnight then heated between50-70° C. for 2 h. The reaction was cooled to room temperature,concentrated under reduced pressure and the residue washed with aqueoussodium bicarbonate solution (1×50 mL). The solid residue was suspendedin a mixture of ethyl acetate (100 mL) and dichloromethane (100 mL) andsonicated. The solids were collected by filtration to give the titlecompound (1.39 g) as an off-white solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 12.28 (s, 1H), 9.76 (s,1H), 7.60 (d, 2H), 7.53 (d, 2H), 4.67-4.59 (m, 1H), 3.72-3.64 (m, 1H),3.68 (s, 3H), 3.55-3.53 (m, 2H), 3.11 (t, 1H), 2.84 (s, 3H), 2.68-2.64(m, 2H), 2.41-2.38 (m, 1H), 2.02-1.93 (m, 1H), 1.79-1.66 (m, 3H), 1.37(s, 2.5H), 1.37-1.12 (m, 3H), 1.11 (s, 6.5H).

ESI MS m/z 582 (M+H)⁺

Example 66 methyl[4-(2-{(2S,4R)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatehydrochloride

To a solution of the compound prepared in Example 64 (0.447 g, 0.811mmol) in 1,4-dioxane was added 4 M HCl in 1,4-dioxane (5.0 mL) and thereaction mixture stirred at room temperature for 1 h. The mixture wasconcentrated and the residue was purified by trituration with 1:1dichloromethane/hexanes to afford the title compound (0.46 g, 99%) as alight brown solid.

¹H NMR (400 MHz, DMSO-d₆) δ 10.20 (br. s, 1H), 9.81 (br. s, 1H), 7.89(br. s, 1H), 7.81 (d, 2H), 7.56 (d, 2H), 4.93-4.89 (m, 1H), 3.68 (s,3H), 3.65-3.52 (m, 2H), 3.50-3.46 (m, 3H), 3.27-3.24 (m, 1H), 2.86 (s,3H), 2.72-2.69 (m, 2H), 2.68-2.65 (m, 1H), 2.22-2.19 (m, 2H), 1.81-1.79(m, 2H), 1.29-1.27 (m, 2H).

Example 67 methyl[4-(4-chloro-2-{(2S,4R)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatehydrochloride

The compound prepared in Example 65 was treated following the proceduredescribed in Example 66 to give the title compound as a light yellowsolid.

¹H NMR (400 MHz, DMSO-d₆) δ 13.50 (br. s, 1H), 9.95 (br. s, 1H), 9.82(s, 1H), 9.28 (br. s, 1H), 7.68 (d, 2H), 7.58 (d, 2H), 4.73-4.69 (m,1H), 3.68 (s, 3H), 3.59-3.55 (m, 3H), 3.56-3.44 (m, 2H), 3.06-3.01 (m,1H), 2.85 (s, 3H), 2.67 (t, 1H), 2.20-2.18 (m, 1H), 1.96-1.90 (m, 1H),1.78-1.76 (m, 2H), 1.46-1.43 (m, 1H), 1.39-1.32 (m, 2H).

Example 68bis(2-methyl-2-propanyl){[4-({(2S,4R)-2-(5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)-1-piperidinyl]methylylidene}biscarbamate

To a solution of the compound prepared in Example 66 (0.11 g, 0.25 mmol)in dichloromethane (5.0 mL) and N,N-dimethylformamide (2.0 mL) was added1-hydroxy-7-azabenzotriazole (0.034 g, 0.25 mmol), followed by N-methylmorpholine (0.075 g, 0.74 mmol),l-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.095 g,0.50 mmol) and the compound prepared in Example 7 (0.10 g, 0.27 mmol).The reaction was stirred at room temperature under nitrogen overnightwhereupon it was concentrated under reduced pressure. The resultantresidue was diluted with dichloromethane (20 mL), washed with brine (10mL), dried and concentrated. Purification by flash chromatography(silica gel, 12 g, 0-5% methanol/dichloromethane) and trituration frommethanol and methyl tert-butyl ether afforded the title compound (0.067g, 38%) as a yellow solid.

¹H NMR (300 MHz, CDCl₃, rotamers present) δ 10.84 (s, 0.3H), 10.54 (s,0.7H), 10.16 (s, 1H), 7.72-7.65 (d, 1.4H), 7.46-7.31 (m, 2.6H),7.20-7.08 (m, 1H), 6.65-6.50 (m, 1H), 5.30-5.16 (t, 1H), 4.28-4.10 (m,2H), 3.93-3.81 (m, 3H), 3.78 (s, 3H), 3.35-3.15 (m, 1H), 3.10-2.98 (m,2H), 2.92-2.87 (m, 0.7H), 2.85-2.77 (m, 0.3H), 2.78 (s, 3H), 2.71-2.57(m, 31H), 2.51-2.45 (m, 1H), 2.18-2.10 (m, 0.6H), 2.08-1.90 (m, 2.41),1.81-1.65 (m, 4H), 1.49 (s, 18H), 1.50-1.32 (m, 3H).

ESI MS m/z 801 (M+H)⁺

Example 69 methyl[4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatedihydrochloride

To a solution of the compound prepared in Example 68 (0.067 g, 0.084mmol) in dichloromethane (2 mL) at 0° C. was added trifluoroacetic acid(0.09 mL, 1.2 mmol). The reaction was warmed to room temperature andstirred for 4 h whereupon the solvent and excess trifluoroacetic acidwas removed in vacuo. The resulting residue was dissolved in methanol (5mL) and 6 M hydrochloric acid (0.70 mL) was added. The mixture wasconcentrated to dryness and the process repeated twice. The residue wastriturated with methanol and methyl tert-butyl ether, the resultingsolids dissolved in water and lyophilized to afford the title compound(0.050 g, 96%) as a yellow solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 15.04 (s, 1H), 14.66 (s,1H), 9.88 (s, 1H), 7.88 (s, 1H), 7.84-7.79 (d, 2H), 7.60-7.54 (d, 2H),7.33 (app. s, 3H), 5.06-4.99 (m, 1H), 4.05-3.90 (m, 1H), 3.88-3.75 (m,3H), 3.68 (s, 3H), 3.63-3.40 (m, 3H), 3.15-3.00 (m, 2H), 2.85 (s, 3H),2.78-2.60 (m, 2H), 2.25-2.10 (m, 1H), 2.05-1.92 (m, 1H), 1.91-1.65 (m,4H), 1.52-1.39 (m, 2H), 1.39-1.10 (m, 3H).

ESI MS m/z 601 (M+H)⁺

Example 70bis(2-methyl-2-propanyl){[4-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)-1-piperidinyl]methylylidene}biscarbomate

The compound prepared in Example 67 was treated with the compoundprepared in Example 7 following the procedure described in Example 68 togive the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 10.91 (s, 1H), 10.16 (s,1H), 7.56 (d, 2H), 7.45 (d, 2H), 6.64 (s, 1H), 5.15 (t, 1H), 4.25-4.11(m, 2H), 3.88-3.85 (m, 3H), 3.79 (s, 3H), 3.16 (t, 1H), 3.08-2.99 (m,2H), 2.77 (s, 3H), 2.70-2.60 (m, 4H), 2.51-2.44 (m, 1H), 2.04-1.92 (m,3H), 1.81-1.70 (m, 4H), 1.49 (s, 9H), 1.47 (s, 9H), 1.59-1.40 (m, 3H).

ESI MS m/z 835 (M+H)⁺

Example 71 methyl[4-(2-{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatedihydrochloride

The compound prepared in Example 70 was treated following the proceduredescribed in Example 69 to give the title compound as a white solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 13.05 (s, 0.4H), 12.68 (s,0.6H), 9.80 (s, 0.4H), 9.79 (s, 0.6H), 7.65 (d, 0.8H), 7.60 (d, 1.2H),7.56-7.53 (m, 2H), 7.31 (app. s, 2.4H), 7.26 (app. s, 1.6H), 5.12-5.10(m, 0.4H), 4.84-4.80 (m, 0.6H), 3.99-3.95 (m, 1H), 3.84-3.79 (m, 1H),3.68 (s, 3H), 3.58-3.53 (m, 2.4H), 3.40 (t, 0.6H), 3.09-3.07 (m, 2H),2.84 (s, 3H), 2.72-2.64 (m, 3H), 2.49-2.43 (m, 2H), 2.10-1.97 (m, 0.6H),1.87-1.65 (m, 5.4H), 1.52-1.19 (m, 5H).

ESI MS m/z 635 (M+H)⁺

Example 72 2-methyl-2-propanyl{imino[4-({(2S,4R)-2-(5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)phenyl]methyl}carbamate

The compound prepared in Example 66 was treated with the compoundprepared in Example 18 following the procedure described in Example 8 togive the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 7.90 (d, 21H), 7.71 (s, 1H),7.65-7.52 (d, 2H), 7.44-7.35 (d, 2H), 7.22-7.15 (m, 3H), 6.57 (s, 1H),5.42-5.32 (t, 1H), 3.90-3.85 (m, 1H), 3.78 (s, 3H), 3.64-3.52 (m, 1H),3.51-3.45 (d, 1H), 3.38-3.22 (t, 1H), 2.77 (s, 3H), 2.69-2.54 (m, 2H),2.10-1.88 (m, 2H), 1.53 (s, 9H), 1.43-1.38 (m, 1H), 1.35-1.20 (m, 4H),0.85-0.90 (m, 1H).

ESI MS m/z 694 (M+H)⁺

Example 73 methyl[4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatedihydrochloride

The compound prepared in Example 72 was treated following the proceduredescribed in Example 69 to give the title compound as an off-whitesolid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 15.25 (s, 1H), 14.87 (s,1H), 9.88 (s, 1H), 9.46-9.38 (m, 2H), 9.15-9.05 (m, 2H), 8.05-7.75 (m,6H), 7.65-7.50 (d, 2H), 5.35-5.22 (m, 1H), 4.10-3.89 (m, 1H), 3.68 (s,3H), 2.83 (s, 3H), 2.70-2.55 (m, 3H), 2.40-2.34 (m, 1H), 2.25-2.15 (m,1H), 2.09-1.90 (m, 2H), 1.85-1.75 (m, 1H), 1.60-1.50 (m, 1H), 1.50-1.35(m, 2H), 1.34-1.15 (m, 1H), 0.85-0.90 (m, 1H).

ESI MS m/z 594 (M+H)⁺

Example 74 2-methyl-2-propanyl{[4-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)phenyl](imino)methyl}carbamate

The compound prepared in Example 67 was treated with the compoundprepared in Example 18 following the procedure described in Example 8 togive the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 10.89-10.88 (m, 1H), 7.89(d, 2H), 7.57-7.54 (m, 4H), 7.40 (d, 2H), 6.67 (s, 1H), 5.33 (t, 1H),3.85-3.82 (m, 1H), 3.78 (s, 3H), 3.77-3.75 (m, 1H), 3.59 (dd, 1H), 3.29(t, 1H), 2.77 (s, 3H), 2.76-2.56 (m, 4H), 1.95-1.94 (m, 2H), 1.59-1.53(m, 1H), 1.55 (s, 9H), 1.42-1.25 (m, 3H).

ESI MS m/z 728 (M+H)⁺

Example 75 methyl[4-(2-{(2S,4R)-1-(4-carbamimidoylbenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatedihydrochloride

The compound prepared in Example 74 was treated following the proceduredescribed in Example 69 to give the title compound as a white solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 12.80 (s, 0.8H), 12.25 (s,0.2H), 9.78 (s, 1H), 9.41 (s, 1.6H), 9.25 (s, 0.4H), 9.09 (s, 1.6H),8.96 (s, 0.4H), 7.88 (d, 1.6H), 7.83 (d, 1.6H), 7.76 (d, 0.4H), 7.63 (d,1.6H), 7.54 (d, 1.6H), 7.49 (d, 0.4H), 7.39-7.34 (m, 0.8H), 5.08 (dd,0.8H), 4.92 (t, 0.2H), 3.67 (s, 3H), 3.67-3.64 (m, 1H), 3.51-3.47 (m,3H), 2.82 (s, 3H), 2.67-2.61 (m, 2H), 2.07-1.78 (m, 4H), 1.53-1.51 (m,1H), 1.39-1.36 (m, 1H), 1.23-1.16 (m, 2H).

ESI MS m/z 628 (M+H)⁺

Example 76 methyl[4-(2-{(2R,4S)-1-[4-(N′,N″-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidamido)benzoyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate

The compound prepared in Example 66 was treated with the compoundprepared in Example 10 following the procedure described in Example 8 togive the title compound as a light brown solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 9.53 (s, 0.5H), 9.38 (s,0.5H), 7.73 (d, 1H), 7.66 (d, 11H), 7.54 (d, 1H), 7.42-7.39 (m, 2H),7.34-7.29 (m, 0.5H), 7.21-7.18 (m, 1.5H), 6.99-6.96 (m, 1H), 6.60-6.58(m, 1H), 5.40-5.38 (m, 0.5H), 4.51-4.29 (m, 0.5H), 3.86-3.83 (m, 2H),3.80 (s, 3H), 3.50-3.48 (m, 1H), 2.95-2.93 (m, 0.5H), 2.79-2.76 (m,2.5H), 2.71-2.58 (m, 2H), 2.01-1.99 (m, 2H), 1.63-1.59 (m, 6H),1.50-1.47 (m, 2H), 1.32-1.24 (m, 18H).

Example 77 methyl[4-(2-{(2S,4R)-1-(4-carbamimidamidobenzoyl)-4-[1-(methylsulfonyl)-4-piperidiny]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 76 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.88 (d, 2H), 7.73 (s, 1H),7.66 (d, 2H), 7.60 (d, 211H), 7.40 (d, 2H), 5.40 (q, 1H), 3.86-3.35 (m,8H), 2.80 (s, 3H), 2.72-2.69 (m, 3H), 2.21-2.14 (m, 1H), 2.00-1.97 (m,2H), 1.71-1.68 (m, 1H), 1.67-1.32 (m, 3H).

APCI MS m/z 609 (M+H)⁺

Example 78 methyl[4-(2-{(2R,4S)-1-[4-(N′,N″-bis{[(2-methyl-2-propanyl)oxy]carbonyl}carbamimidamido)benzoyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamate

The compound prepared in Example 67 was treated with the compoundprepared in Example 10 following the procedure described in Example 8 togive the title compound as a light brown solid.

¹H NMR (300 MHz, CDCl₃, rotamers present) δ 10.90 (br. s, 1H), 7.59 (d,2H), 7.52 (d, 2H), 7.42 (d, 2H), 7.19 (d, 2H), 6.73 (s, 1H), 5.41-5.30(m, 1H), 3.91-3.79 (m, 4H), 3.78 (s, 3H), 3.36 (t, 1H), 3.21-3.06 (m,1H), 2.77 (s, 3H), 2.76-2.51 (m, 4H), 2.02-1.83 (m, 2H), 1.81-1.78 (m,2H), 1.41-1.27 (m, 18H).

Example 79 methyl[4-(2-{(2S,4R)-1-(4-carbamimidamidobenzoyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 78 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.74 (d, 2H), 7.64 (d, 2H),7.54 (d, 2H), 7.37 (d, 2H), 5.19 (dd, 1H), 3.74 (s, 3H), 3.71-3.65 (m,4H), 2.79 (s, 3H), 2.72-3.69 (m, 5H), 2.15-2.01 (m, 1H), 1.98-1.87 (m,3H), 1.58-1.49 (m, 1H).

ESI MS m/z 643 (M+H)⁺

Example 80 2-methyl-2-propanyl{[cis-4-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)cyclohexyl]methyl}carbamate

Following the procedure described in Example 8, the compound prepared inExample 67 was treated withcis-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexanecarboxylicacid to give the title compound as a white solid.

¹H NMR (500 MHz, CDCl₃, rotamers present. One exchangeable proton wasnot observed.) δ 11.12 (s, 1H), 7.65-7.35 (m, 4H), 6.80-6.64 (m, 1H),5.80-5.06 (m, 1H), 4.64-4.52 (m, 1H), 3.88-3.81 (m, 3H), 3.79 (s, 3H),3.09-2.97 (m, 2H), 2.78 (s, 3H), 2.67-2.56 (m, 3H), 2.56-2.44 (m, 2H),2.05-1.92 (m, 2H), 1.86-1.61 (m, 5H), 1.60-1.49 (m, 6H), 1.43 (s, 9H),1.32-1.23 (m, 2H).

ESI MS m/z 721 (M+H)⁺

Example 81 methyl[4-(2-{(2S,4R)-1-{[cis-4-(aminomethyl)cyclohexyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatedihydrochloride

The compound prepared in Example 80 was treated following the proceduredescribed in Example 69 to give the title compound as a white solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 12.84 (s, 0.3H), 12.47 (s,0.7H), 9.81-9.76 (m, 1H), 7.76-7.48 (m, 7H), 5.00 (t, 0.3H), 4.81 (t,0.7H), 4.03-3.70 (m, 2H), 3.68 (s, 3H), 3.59-3.51 (m, 2H), 3.39-3.30 (m,1H), 2.85 (s, 3H), 2.78-2.59 (m, 4H), 2.44-2.34 (m, 2H), 2.07-1.96 (m,1H), 1.83-1.72 (m, 4H), 1.72-1.63 (m, 1H), 1.63-1.22 (m, 9H).

ESI MS m/z 621 (M+H)⁺

Example 82 2-methyl-2-propanyl{(1S)-1-[trans-4-({(2S,4R)-2-(5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)cyclohexyl]ethyl}carbamate

Following the procedure described in Example 8, the compound prepared inExample 66 was treated withtrans-4-[(1S)-1-({[(2-methyl-2-propanyl)oxy]carbonyl}amino)ethyl]cyclohexanecarboxylicacid to give the title compound as a light brown solid.

¹H NMR (400 MHz, CDCl₃, rotamers present) δ 7.52 (br s, 2H), 7.37 (d,2H), 7.15 (s, 1H), 6.63 (br s, 1H), 5.20 (t, 1H), 4.44-4.41 (m, 1H),3.92-3.80 (m, 3H), 3.78 (s, 3H), 3.60-3.49 (m, 1H), 3.22-3.10 (m, 1H),2.79 (s, 3H), 2.71-2.59 (m, 2H), 2.55-2.48 (m, 1H), 2.41-2.29 (m, 1H),2.12-2.09 (m, 1H), 2.08-1.98 (m, 1H), 1.91-1.70 (m, 6H), 1.65-1.49 (m,4H), 1.44 (s, 9H), 1.38-1.32 (m, 2H), 1.09 (d, 3H), 1.04-1.03 (m, 1H).

Example 83 methyl[4-(2-{(2S,4R)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonal)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 82 was treated following the proceduredescribed in Example 9 to give the title compound as a white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.67 (s, 1H), 7.60-7.51 (m,4H), 5.15-5.05 (m, 1H), 4.11-4.01 (m, 1H), 3.80-3.70 (m, 5H), 3.61-3.49(m, 1H), 3.19-3.09 (m, 1H), 2.82 (s, 3H), 2.80-2.65 (m, 3H), 2.62-3.50(m, 1H), 2.35-2.20 (m, 1H), 1.99-1.80 (m, 7H), 1.61-1.39 (m, 6H), 1.26(d, 3H), 1.24-1.14 (m, 2H).

APCI MS m/z 601 (M+H)⁺

Example 84 methyl[4-(4-chloro-2-{(2S,4R)-1-({4-[(methylamino)methyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

Following the procedure described in Example 8, the compound prepared inExample 67 was treated withtrans-4-[(N-methyl-{[(2-methyl-2-propanyl)oxy]carbonyl})amino)methyl]cyclohexanecarboxylicacid to give the crude amide. After this, the crude amide was treatedfollowing the procedure described in Example 9 to give the titlecompound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.67-7.57 (m, 4H), 5.10 (dd,1H), 4.06-4.00 (m, 1H), 3.75-3.68 (m, 5H), 3.60 (t, 1H), 2.87-2.81 (m,6H), 2.74-2.66 (m, 8H), 2.29-2.26 (m, 2H), 2.23-1.67 (m, 5H), 1.49-1.28(m, 4H), 1.17-0.60 (m, 3H).

ESI MS m/z 635 (M+H)⁺

Example 85 2-methyl-2-propanyl{(1S)-1-[trans-4-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)cyclohexyl]ethyl}carbamate

Following the procedure described in Example 8, the compound prepared inExample 67 was treated withtrans-4-[(1S)-1-({[(2-methyl-2-propanyl)oxy]carbonyl}amino)ethyl]cyclohexanecarboxylicacid to give the title compound as a light brown solid.

¹H NMR (300 MHz, CDCl₃, rotamers present) δ 11.0 (br. s, 1H), 7.57 (d,2H), 7.45 (d, 2H), 6.68 (s, 1H), 5.17 (t, 1H), 4.34 (d, 1H), 3.87-3.83(m, 3H), 3.79 (s, 3H), 3.52-3.49 (m, 1H), 3.16 (t, 1H), 2.88 (s, 3H),2.68-2.59 (m, 3H), 2.50-2.47 (m, 1H), 2.30-2.28 (m, 1H), 2.00-1.87 (m,3H), 1.85-1.73 (m, 5H), 1.48-1.46 (m, 1H), 1.45 (s, 9H), 1.25-1.23 (m,4H), 1.09 (d, 3H), 1.06-1.03 (m, 1H).

Example 86 methyl[4-(2-{(2S,4R)-1-({trans-4-[(1S)-1-aminoethyl]cyclohexyl}carbonyl)-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 85 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (400 MHz, CD₃OD, rotamers present) δ 7.62 (d, 2H), 7.53 (d, 2H),5.08-5.06 (m, 1H), 4.12-4.04 (m, 1H), 3.75 (app. s, 5H), 3.54-3.49 (m,1H), 3.19-3.03 (m, 1H), 2.82 (s, 3H), 2.73-2.70 (m, 3H), 2.56-2.53 (m,1H), 2.35-2.24 (m, 1H), 2.01-1.81 (m, 6H), 1.63-1.31 (m, 7H), 1.26 (d,3H), 1.21-1.19 (m, 2H).

APCI MS m/z 635 (M+H)⁺

Example 87bis(2-methyl-2-propanyl){[4-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl-1-piperazinyl]methylylidene}biscarbamate

The compound prepared in Example 67 was treated with the compoundprepared in Example 28 following the procedure described in Example 29to give the title compound as a white solid.

¹H NMR (400 MHz, DMSO-d₆, rotamers present) δ 12.3 (s, 1H), 9.75 (s,1H), 9.58 (s, 1H), 7.56 (d, 2H), 7.54 (d, 2H), 5.32-5.31 (m, 1H), 4.93(t, 1H), 3.67 (s, 3H), 3.54-3.45 (m, 4H), 3.41-3.31 (m, 2H), 3.17-3.16(m, 3H), 2.84 (s, 3H), 2.67-2.66 (m, 3H), 2.03-1.89 (m, 4H), 1.81-1.75(m, 3H), 1.41 (s, 9H), 1.36 (s, 9H), 1.31-1.29 (m, 2H).

Example 88 methyl[4-(2-{(2S,4R)-1-[(4-carbamimidoyl-1-piperazinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 87 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.62 (d, 2H), 7.71 (d, 2H),5.06-5.05 (m, 1H), 3.75 (s, 3H), 3.71-3.65 (m, 2H), 3.64-3.63 (m, 4H),3.57-3.54 (m, 5H), 2.81 (s, 3H), 2.73 (t, 2H), 2.52-2.48 (m, 1H),1.99-1.89 (m, 4H), 1.45-1.36 (m, 4H).

ESI MS, m/z 636 (M+H)⁺

Example 89 2-methyl-2-propanyl{[1-({(2S,4R)-2-(4-chloro-5-{4-[(methoxycarbonyl)amino]phenyl}-1H-imidazol-2-yl)-4-[1-(methylsulfonyl)-4-piperidinyl]-1-pyrrolidinyl}carbonyl)-4-piperidinyl]methyl}carbamate

Following the procedure described in Example 29, the compound preparedin Example 67 was treated with4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]piperidine to givethe title compound as a white solid.

¹H NMR (400 MHz, CDCl₃ rotamers present. One exchangeable proton was notobserved.) δ 11.0 (s, 1H), 7.52-7.31 (m, 4H), 6.85 (s, 1H), 5.18-5.11(m, 1H), 4.64-4.35 (m, 1H), 3.82-3.68 (m, 7H), 3.51-3.47 (m, 1H), 3.25(t, 1H), 3.28-3.25 (m, 1H), 2.94-2.88 (m, 2H), 2.66 (s, 4H), 2.64-2.58(m, 3H), 2.42-2.31 (m, 2H), 1.97-1.91 (m, 3H), 1.74-1.70 (m, 12H),1.39-1.28 (m, 3H).

Example 90 methyl[4-(2-{(2S,4R)-1-{[4-(aminomethyl-1-piperidinyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

The compound prepared in Example 89 was treated following the proceduredescribed in Example 9 to give the title compound as an off-white solid.

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.61 (d, 2H), 7.53 (d, 2H),5.05 (dd, 1H), 3.95-3.90 (m, 1H), 3.88-3.81 (m, 1H), 3.76-3.68 (m, 4H),3.57 (t, 1H), 3.50-3.48 (m, 1H), 2.92-2.89 (m, 1H), 2.85-2.73 (m, 6H),2.70-2.48 (m, 3H), 2.47-2.45 (m, 1H), 2.03-2.09 (m, 1H), 1.88-1.70 (m,4H), 1.46-1.14 (m, 7H).

ESI MS m/z 622 (M+H)⁺

Example 914-[({(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid

Allyl4-[({(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoatewas prepared by following the procedures described in Examples 5, 9 and8 starting from the compound prepared in Example 63. (Note: in the stepcorresponding to Example 5, allyl 4-aminobenzoate was used in place oftert-butyl 4-aminobenzoate. In the step corresponding to Example 8,(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid was used inplace of1-(N,N′-bis(tert-butoxycarbonyl)carbamimidoyl)piperidine-4-carboxylicacid). After that, to a solution of the crude allyl ester (48 mg) inN,N-dimethylformamide (1 mL) was addedtetrakis(triphenylphosphine)palladium(0) (2 mg) and1,3-dimethylbarbituric acid (4 mg) and the reaction stirred overnight atroom temperature. Further tetrakis(triphenylphosphine)palladium(0) (3mg) was added at this juncture and stirring continued at roomtemperature for 7 h. The reaction mixture was left to stand overnight,diluted with a 1:1 mixture of diisopropylether and dichloromethane (1mL) and the resulting precipitate collected by filtration. The yellowpowder obtained was washed with dichloromethane and dried to give thetitle compound having the following physical properties (25 mg).

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 12.69 (br. s, 1H), 10.38(s, 1H), 9.87 (s, 1H), 8.38 (d, 1H), 7.88 (d, 2H), 7.77 (dd, 1H), 7.72(d, 1H), 7.68 (d, 2H), 7.30 (d, 1H), 6.85 (d, 1H), 4.42 (t, 1H), 4.21(dd, 1H), 3.62-3.50 (m, 2H), 3.37-3.25 (obs. m, 1H), 2.86 (s, 3H),2.75-2.62 (m, 2H), 2.45-2.37 (m, 1H), 2.14-2.01 (m, 1H), 1.88-1.77 (m,2H), 1.53 (dd, 1H), 1.43-1.20 (m, 3H).

ESI MS m/z 628 (M+H)⁺

Example 92 3-chloro-4-fluoro-1-methyl-1H-indole-5-carboxylic acid

To a solution of4-fluoro-1-[tris(propan-2-yl)silyl]-1H-indole-5-carboxylic acid (1.86 g)in dichloromethane (18 mL) and N,N-dimethylformamide (7 mL) was addedN-chlorosuccinimide (0.741 g) and the reaction stirred at roomtemperature under nitrogen for 3 h. Dimethylsulfoxide (10 mL) was addedand the reaction stirred at room temperature for 1.5 h. Thedichloromethane was removed in vacuo and the reaction left to stand atroom temperature for 7 days. The reaction was then diluted withdichloromethane (8 mL) and N,N-dimethylformamide (12 mL) and furtherN-chlorosuccinamide (0.518 g) added. The reaction was stirred for 4 h,N-chlorosuccinamide (0.518 g) added and stirring continued over 16hours. Further N-chlorosuccinamide (0.518 g×3) was added and thereaction mixture stirred for a further 24 hours. The reaction mixturewas concentrated in vacuo and the residue partitioned between ethylacetate and water. The aqueous layer was acidified with 1 M hydrochloricacid (pH=1-2) and extracted with ethyl acetate. The combined organiclayers were washed with water, dried and concentrated.

The residue thus obtained was dissolved in N,N-dimethylformamide (15 mL)and the solution cooled to 0° C. Sodium hydride (611 mg, 63% dispersionin oil) was added and the reaction stirred at 0° C. for 10 minutes,methyl iodide (1.33 mL) was then added and the reaction stirred afurther 10 minutes before diluting with N,N-dimethylformamide (15 mL)and stirring for 40 minutes at room temperature. A saturated solution ofaqueous ammonium chloride (20 mL) was added and the reaction partitionedbetween ethyl acetate and water. The aqueous layer was extracted withethyl acetate and the combined organic fractions washed with water,dried and concentrated. The crude product was purified by columnchromatography (dichloromethane, then 20-30% ethylacetate/dichloromethane) to give methyl3-chloro-4-fluoro-1-methyl-1H-indole-5-carboxylate (1.04 g) as a yellowsolid.

The ester thus obtained was dissolved in a mixture of tetrahydrofuran(10 mL), methanol (10 mL) and dichloromethane (3 mL). 2 M sodiumhydroxide (4.31 mL) was added and the mixture stirred at roomtemperature for 3 h. Further 2 M sodium hydroxide (4.31 mL) was addedand the reaction stirred at room temperature for 16 h. The mixture wasconcentrated in vacuo, diluted with water and ethyl acetate added. Theresultant emulsion was acidified with 1 M hydrochloric acid (pH=2-3) andextracted with ethyl acetate. The combined ethyl acetate phases weredried and concentrated to give the title compound (0.274 g) as a paleyellow solid.

ESI MS m/z 228 (M+H)⁺

Example 934-[({(2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid

Allyl4-[({(2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoatewas prepared from the compound prepared in Example 63 following theprocedures described in Example 5, 9 and 8. (Note: in the stepcorresponding to Example 5, allyl 4-aminobenzoate was used in place oftert-butyl 4-aminobenzoate. In the step corresponding to Example 8, theacid prepared in Example 92 was used in place of1-(N,N′-bis(tert-butoxycarbonyl)carbamimidoyl)piperidine-4-carboxylicacid). After that, the crude allyl ester was treated with palladium(0)following the procedure described in Example 91 to give the titlecompound as an off-white solid.

¹H NMR (500 MHz, DMSO-d₆, rotamers present) δ 12.74 (br. s, 1H), 10.50(s, 1H), 7.91 (d, 2H), 7.76 (d, 2H), 7.65 (s, 1H), 7.42 (d, 1H), 7.22(dd, 11H), 4.61 (t, 1H), 3.80 (s, 3H), 3.58-3.38 (m, 3H), 3.27 (t, 1H),2.80 (s, 3H), 2.65-2.55 (m, 3H), 2.09-1.98 (m, 1H), 1.82-1.73 (m, 1H),1.62 (dd, 1H), 1.52-1.45 (m, 1H), 1.40-1.26 (m, 1H), 1.25-1.09 (m, 2H).

ESI MS m/z 605 (M+H)⁺

Example 94 methyl[4-(4-chloro-2-{(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetraol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate

Following the procedure described in Example 68, the compound preparedin Example 67 was treated with(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid to give thetitle compound as a white solid.

¹H NMR (300 MHz, DMSO-d₆, rotamers present) δ 12.80 (br. s, 0.3H), 12.40(br. s, 0.7H), 9.95-9.64 (m, 2H), 8.42-6.58 (m, 8H), 7.24 (d, 0.7H),6.97 (d, 0.3H), 6.80 (d, 0.7H), 6.67 (d, 0.3H), 5.23 (t, 0.3H), 4.86 (t,0.7H), 4.18 (br. t, 0.7H), 3.97 (q, 0.3H), 3.80-2.30 (m, 12H), 2.20-1.61(m, 3H), 1.55-0.96 (m, 3H).

FAB MS m/z 714 (M+H)⁺

Example 95 methyl[4-(4-chloro-2-{2S,4R)-1-[(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate

Following the procedure described in Example 68, the compound preparedin Example 67 was treated with the compound prepared in Example 92 togive the title compound as a white solid.

¹H NMR (300 MHz, DMSO-d₆, rotamers present) δ 12.60 (s, 0.6H), 11.90 (s,0.4H), 9.77 (s, 0.6H), 9.72 (s, 0.4H), 7.81-6.47 (m, 7H), 5.05 (t,0.6H), 4.69 (br. t, 0.4H), 4.14-3.19 (m, 9H), 2.87 (s, 1.8H), 2.71 (s,1.2H), 2.70-2.28 (m, 4H), 2.18-1.66 (m, 4H), 1.59-1.01 (m, 3H).

ESI MS m/z 691 (M+H)⁺

Example 96 methyl[4-(2-{(2S,4R)-1-{[4-(aminomethyl)cyclohexyl]carbonyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatebis(trifluoroacetate)

Following the procedure described in Example 68, the compound preparedin Example 67 was treated withtrans-4-(tert-butoxycarbonylamino)-cyclohexanecarboxylic acid. Afterthis, the crude amide was treated with TFA following the proceduredescribed in Example 9 to give the title compound as a white solid.

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.96-7.22 (m, 4H), 5.27-4.50(m, 1H), 4.18-3.91 (m, 1H), 3.86-3.59 (m, 5H), 3.50 (br. t, 1H),3.00-2.41 (m, 7H), 2.41-2.06 (m, 1H), 2.06-1.69 (m, 8H), 1.69-0.50 (m,9H).

ESI MS m/z 621 (M+H)⁺

Example 97 methyl[4-(2-{(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamatetrihydrochloride

The compound prepared in Example 4 was treated following the proceduresdescribed in Example 64, 6, 8 and 69 to give the title compound as awhite solid. (Note: in the step corresponding to Example 8, the compoundprepared in Example 18 was used)

¹H NMR (300 MHz, CD₃OD, rotamers present) δ 7.91 (d, 2H), 7.89 (d, 2H),7.75 (s, 1H), 7.65 (dd, 2H), 7.60 (dd, 2H), 5.38 (dd, 1H), 3.90-3.60 (m,6H), 2.80 (s, 3H), 2.79-2.60 (m, 3H), 2.35-2.18 (m, 1H), 2.10-1.80 (m,2H), 1.65-1.25 (m, 4H).

ESI MS m/z 595 (M+H)⁺

Example 98 methyl[4-(2-(2S,4S)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl-1H-imidazol-5-yl)phenyl]carbamatetrihydrochloride

The compound prepared in Example 4 was treated following the proceduresdescribed in Example 64, 6, 8 and 69 to give the title compound as awhite solid. (Note: in the step corresponding to Example 8, the compoundprepared in Example 7 was used)

¹H NMR (300 MHz, DMSO-d₆, rotamers present) & 9.90 (s, 1H), 7.94 (s,1H), 7.85 (d, 2H), 7.60 (s, 2H), 7.52-7.31 (m, 3H), 5.21 (t, 1H),4.51-2.20 (m, 24H), 2.05-1.89 (m, 1H), 1.89-1.69 (m, 1H), 1.57-1.25 (m,2H).

ESI MS m/z 602 (M+H)⁺

Example 99 methyl[4-(2-{(2S,4S)-1-(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[4-(methylsulfonyl)-1-piperazinyl-2-pyrrolidinyl}-4-chloro-1H-imidazol-5-yl)phenyl]carbamatetrihydrochloride

The compound prepared in Example 4 was treated following the proceduresdescribed in Example 64, 65, 6, 8 and 69 to give the title compound as awhite solid. (Note: in the step corresponding to Example 8, the compoundprepared in Example 7 was used)

¹H NMR (300 MHz, DMSO-d₆, rotamers present) δ 9.83 (s, 0.5H), 9.84 (s,0.5H), 7.84 (d, 1H), 7.24 (d, 1H), 7.58-7.49 (m, 2H), 7.49-7.18 (m, 4H),5.40 (t, 0.5H), 4.93 (t, 0.5H), 4.42-2.30 (m, 24H), 1.93-0.49 (m, 4H).

ESI MS m/z 636 (M+H)⁺

Example 100(3-chloro-4-fluoro-1-methyl-1H-indol-5-yl)[(2S,4S)-2-(4-chloro-5-phenyl-1H-imidazol-2-yl-4-(4-morpholinyl)-1-pyrrolidinyl]methanonebis(trifluoroacetate)

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 64, 65, 6 and 8 to give the title compoundafter purification by high performance liquid chromatography [5 to 100%mobile phase B (0.1% trifluoroacetic acid in acetonitrile) in mobilephase A (0.1% trifluoroacetic acid in water)] as a white solid. (Note:in the steps corresponding to Examples 3, 64 and 8, morpholine,2-bromo-1-phenylethane-1-one and the compound prepared in Example 92were used respectively)

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 10.92 (app. br. s, 3H), 7.81(d, 1H), 7.72-7.58 (m, 1H), 7.49-7.31 (m, 3H), 7.30-6.99 (m, 3H),6.02-5.62 (m, 1H), 4.65-4.28 (m, 1H), 4.24-3.84 (m, 6H), 3.81-3.61 (m,3H), 3.60-2.89 (m, 6H).

ESI MS m/z 542/544 (M+H)⁺

Example 101(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-1-[(2S,4S)-4-(4-morpholinyl)-2-(5-phenyl-1H-imidazol-2-yl)-1-pyrrolidinyl]-2-propen-1-one

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 64, 6 and 8 to give the title compound as awhite solid. (Note: in the step corresponding to Example 3, 64 and 8,morpholine, 2-bromo-1-phenylethane-1-one and(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid were usedrespectively)

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 8.69 (s, 1H), 7.87 (d, 1H),7.78-7.67 (m, 1H), 7.64 (d, 1H), 7.47 (dd, 1H), 7.43-7.28 (m, 4H),7.26-7.19 (m, 2H), 7.04 (d, 1H), 5.39 (dd, 1H), 4.23 (d, 1H), 3.90-3.71(m, 6H), 3.65 (dd, 1H), 2.97-2.81 (m, 1H), 2.78-2.62 (m, 2H), 2.63-2.54(m, 2H).

ESI MS m/z 531 (M+H)⁺

Example 102(2E)-1-[(2S,4S)-2-(4-chloro-5-phenyl-1H-imidazol-2-yl)-4-(4-morpholinyl)-1-pyrrolidinyl]-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-proven-1-one

The compound prepared in Example 2 was treated following the proceduresdescribed in Examples 3, 4, 64, 65, 6 and 8 to give the title compoundas a white solid. (Note: in the step corresponding to Example 3, 64 and8, morpholine, 2-bromo-1-phenylethane-1-one and(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]acrylic acid were usedrespectively)

¹H NMR (500 MHz, CDCl₃, rotamers present) δ 12.60 (br. s, 1H), 8.85 (s,1H), 7.90 (s, 1H), 7.74-7.60 (m, 2H), 7.57 (dd, 1H), 7.49-7.38 (m, 3H),7.38-7.20 (m, 2H), 7.10 (d, 1H), 5.32-5.21 (m, 1H), 4.25 (d, 1H), 3.96(dd, 1H), 3.87-3.57 (m, 4H), 3.08 (br. s, 1H), 2.90 (dd, 1H), 2.75-2.40(m, 5H).

ESI MS m/z 565/567 (M+H)⁺

Example 103 benzyl 3-acetylbenzoate

To a solution of 3-acetylbenzoic acid (6.00 g, 0.36 mol) in DMF (36 mL)was added Na₂CO₃ (4.26 g, 0.40 mol) at 0° C. and the reaction stirredfor 10 min. After 10 min, benzyl bromide (4.80 mL, 0.40 mol) was addedand the reaction stirred at room temperature for 1 h. The reactionmixture was diluted with EtOAc (100 mL) and washed with H₂O (50 mL) andbrine (50 mL), dried over Na₂SO₄, filtered and concentrated. The residuewas purified by combiflash chromatography (silica gel, 40 g, 2-5% ethylacetate/hexanes) to afford the title compound (7.73 g, 84%) as a whitesolid.

¹H NMR (300 MHz, CDCl₃) δ 8.61 (dd, 1H), 8.26 (ddd, 1H), 8.15 (ddd, 1H),7.54 (t, 1H), 7.45-7.37 (m, 5H), 5.40 (s, 2H), 2.63 (s, 3H).

Example 104 benzyl 3-(2-bromoacetyl)benzoate

To a solution of compound prepared in Example 103 (8.40 g, 0.33 mol) inTHF (160 mL) was added Phenyl Trimethylammonium Tribromide (12.40 g,0.33 mol) and the reaction mixture stirred at room temperature for 1 h.The reaction mixture was diluted with EtOAc (100 mL) and washed with H₂O(50 mL) and brine (50 mL), dried over Na₂SO₄, filtered and concentrated.The residue was purified by combiflash chromatography (silica gel, 40 g,2-3% ethyl acetate/hexanes) to afford the title compound (7.99 g, 55%)as a white solid.

¹H NMR (300 MHz, CDCl₃) δ 8.64 (dd, 1H), 8.31 (ddd, 1H), 8.18 (ddd, 1H),7.58 (t, 1H), 7.47-7.36 (m, 5H), 5.40 (s, 2H), 4.47 (s, 2H).

Example 1053-[2-[(2S,4R)-1-(1-carbamimidoylpiperidine-4-carbonyl)-4-(1-methylsulfonyl-4-piperidyl)pyrrolidin-2-yl]-1H-imidazol-5-yl]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 63 was treated following the proceduresdescribed in Examples 64, 66, 68, 6 and 9 to give the title compound asa white solid. (Note: in the steps corresponding to Examples 64, thecompound prepared in Example 104 was used)

¹H NMR (300 MHz, D₂O) δ 8.19 (s, 1H), 7.98 (d, 1H), 7.83 (d, 1H), 7.61(s, 1H), 7.61 (t, 1H), 5.19-5.16 (m, 1H), 4.11 (t, 1H), 3.85-3.81 (m,2H), 3.70-3.57 (m, 3H), 3.20-3.11 (m, 2H), 3.00-2.98 (m, 4H), 2.84-2.71(m, 3H), 2.32-2.30 (m, 1H), 1.92-1.80 (m, 5H), 1.66-1.60 (m, 3H),1.48-1.43 (m, 2H).

ESI MS m/z 570 [C₂₇H₃₇N₇O₅S−H]⁻

Example 1063-[2-[(2S,4R)-1-[4-(aminomethyl)cyclohexanecarbonyl]-4-(1-methylsulfonyl-4-piperidyl)pyrrolidin-2-yl]-1H-imidazol-5-yl]benzoicacid bis(trifluoroacetate)

The compound prepared in Example 63 was treated following the proceduresdescribed in Examples 64, 66, 68, 6 and 9 to give the title compound asa white solid. (Note: in the steps corresponding to Examples 64 and 68,the compound prepared in Example 104 andtrans-4-[({[(2-methyl-2-propanyl)oxy]carbonyl}amino)methyl]cyclohexanecarboxylicacid were used respectively)

¹H NMR (400 MHz, DMSO-d₆) 8.35 (s, 1H), 8.15 (s, 1H), 8.03-7.97 (m, 2H),7.75 (br s, 3H), 7.65 (t, 1H), 5.03-4.99 (m, 1H), 3.95 (t, 1H),3.62-3.52 (m, 2H), 3.46 (t, 1H), 2.85 (s, 3H), 2.72-2.70 (m, 4H),2.58-2.42 (m, 1H), 2.29-2.12 (m, 1H), 1.89-1.66 (m, 7H), 1.54-0.98 (m,9H).

APCI MS m/z 558 [C₂₈H₃₉N₅O₅S+H]⁺

Example 107 methylN-[4-[2-[(2S,4R)-1-[4-(aminomethyl)cyclohexanecarbonyl]-4-(1-methylsulfonyl-4-piperidyl)pyrrolidin-2-yl]-1H-imidazol-5-yl]phenyl]carbamatebishydrochloride

Following the procedure described in Example 68, the compound preparedin Example 66 was treated withtrans-4-(tert-butoxycarbonylamino)-cyclohexanecarboxylic acid. Afterthis, the crude amide was treated following the procedure described inExample 69 to give the title compound as a white solid.

¹NMR (400 MHz, DMSO-d₆, rotamers present) δ 12.04 (br. s, 0.4H), 9.93(br s, 0.6H), 7.84-7.78 (m, 7H), 7.57-7.52 (m, 2H), 5.11-5.00 (m,0.62H), 3.93-3.86 (m, 0.69H), 3.71-3.68 (m, 1H), 3.68 (s, 3H), 3.61-3.53(m, 2H), 3.52-3.44 (m, 1H), 2.85 (s, 3H), 2.69-2.60 (m, 4H), 2.18-2.09(m, 1H), 1.95-1.89 (m, 2H), 1.82-1.74 (m, 5H), 1.55-1.42 (m, 2H),1.36-1.18 (m, 5H), 1.06-0.98 (m, 3H).

ESI MS m/z 585 [C₂₉H₄₂N₆O₅S−H]⁻

Example 108 benzyl N-[5-(2-bromoacetyl)-2-pyridyl]carbamate

Benzyl N-(5-acetyl-2-pyridyl)carbamate was treated following theprocedures described in Example 103 to give the title compound.

¹H NMR (300 MHz, DMSO-d₆, rotamers present) δ 10.93-10.68 (m, 1H),8.99-8.78 (m, 1H), 8.41-8.19 (m, 1H), 8.05-7.89 (m, 1H), 7.48-7.25 (m,5H), 5.21 (s, 2H), 4.93-4.83 (m, 1.5H), 4.77-4.67 (m, 0.5H).

Example 1094-[(2S,4R)-2-[5-(6-amino-3-pyridyl)-1H-imidazol-2-yl]-4-(1-methylsulfonyl-4-piperidyl)pyrrolidine-1-carbonyl]biperidine-1-carboxamidinetri(trifluoroacetate)

The compound prepared in Example 63 was treated following the proceduresdescribed in Examples 64, 66, 68 and 9 to give the title compound as awhite semi-solid. (Note: in the steps corresponding to Examples 64, thecompound prepared in Example 108 was used)

¹H NMR (400 MHz, D₂O) δ 8.07 (s, 1H), 8.04 (d, 1H), 7.57 (s, 1H), 7.08(d, 1H), 5.12 (t, 1H), 4.07 (t, 1H), 3.80 (d, 2H), 3.64 (d, 2H), 3.53(t, 1H), 3.10 (t, 2H), 2.99-2.23 (m, 1H), 2.91 (s, 3H), 2.82-2.62 (m,3H), 2.33-2.19 (m, 11H), 1.91-1.73 (m, 5H), 1.60-1.44 (m, 3H), 1.44-1.27(m, 2H).

ESI MS m/z 544 [C₂₅H₃₇N₉O₃S+H]⁺

Example 1104-[{(2S,4R)-1-[(1-carbamimidoyl-4-piperidinyl)carbonyl]-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid hydrochloride

The compound prepared in Example 63 was treated following the proceduresdescribed in Examples 5, 66, 68 and 69 to give the title compound as awhite powder.

¹H NMR (CD₃OD, 400 MHz) δ 7.99 (d, 2H), 7.58 (d, 2H), 4.47-4.41 (m, 1H),4.02 (t, 1H), 3.90-3.79 (m, 2H), 3.69-3.61 (m, 2H), 3.36 (t, 1H),3.23-3.08 (m, 2H), 2.94 (s, 4H), 2.82-2.69 (m, 2H), 2.61-2.49 (m, 1H),2.09-2.22 (m, 11H), 1.98-1.74 (m, 4H), 1.72-1.56 (m, 3H) 1.51-1.25 (m,3H).

ESI MS m/z 549 [C₂₅H₃₆N₆O₆S+H]⁺

Example 1114-[({(2S,4R)-1-(4-carbamimidoylbenzoyl-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid hydrochloride

The compound prepared in Example 63 was treated following the proceduresdescribed in Examples 5, 66, 68 and 69 to give the title compound as awhite solid. (Note: in the steps corresponding to Examples 68, thecompound prepared in Example 18 was used)

¹H NMR (CD₃OD, 400 MHz) δ 7.98 (d, 2H), 7.91-7.85 (m, 4H), 7.75 (d, 2H),4.77-4.69 (m, 1H), 3.77-3.60 (m, 3H), 3.51 (t, 1H), 2.79 (s, 3H),2.75-2.60 (m, 3H), 2.17-2.07 (m, 1H), 1.92-1.85 (m, 1H), 1.84-1.73 (m,1H), 1.64-1.56 (m, 1H), 1.49-1.22 (m, 3H).

ESI MS m/z 542 [C₂₆H₃₁N₅O₆S+H]⁺

Example 112 benzyl4-[({(2S,4S)-1-(4-{N′-[(benzyloxy)carbonyl]carbamimidoyl}benzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

The compound prepared in Example 19 was treated following the proceduresdescribed in Examples 69, 2 and 22 to give the title compound as a whitepowder.

¹H NMR (300 MHz, DMSO-d₆) δ 10.49 (s, 1H), 9.17 (br. s, 2H), 8.03 (d,2H), 7.95 (d, 2H), 7.76 (d, 2H), 7.66 (d, 2H), 7.49-7.24 (m, 10H), 5.31(s, 2H), 5.10 (s, 2H), 4.62 (dd, 2H), 3.69-3.59 (m, 1H), 3.53 (t, 1H),3.14-2.88 (m, 5H), 2.83 (s, 3H), 2.64-2.31 (m, 4H), 1.85-1.63 (m, 1H).

Example 113 ethyl4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoatebis(trifluoroacetate)

To a stirred solution of compound prepared in Example 20 (500 mg) inethanol (25 mL) was added concentrated sulfuric acid (20 drops) at 0° C.The resulting mixture was stirred at 60° C. for 44 h whereupon themixture was concentrated in vacuo. The resulting residue was purified byPrep-HPLC (0.1% TFA containing CH₃CN—H₂O gradient) to provide the titlecompound (307 mg).

¹H NMR (300 MHz, D₂O, rotamers present) δ 8.07 (d, 1H), 7.97-7.93 (m,2H), 7.86-7.77 (m, 2H), 7.69-7.65 (m, 2H), 7.22 (m, 1H), 4.90-4.35 (m,3H), 4.12-3.75 (m, 3H), 3.60-3.13 (m, 8H), 3.09-2.97 (m, 4H), 2.31 (m,1H), 1.44-1.34 (m, 3H).

FAB MS m/z 571 (M+H)⁺

Example 114 benzyl4-[({(2S,4S)-1-(4-cyanobenzoyl)-4-[4-(methasulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

The compound prepared in Example 4 was treated following the proceduresdescribed in Examples 5, 6 and 8 to give the title compound having thefollowing physical properties. (Note: in the steps corresponding toExamples 5 and 8, benzyl 4-aminobenzoate and 4-cyanobenzoic acid wereused respectively) TLC: Rf 0.57 (5% methanol in ethyl acetate)

Example 115 benzyl4-[({(2S,4S)-1-[4-(N′-hydroxycarbamimidoyl)benzoyl]-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoate

To a stirred solution of compound prepared in Example 114 (1.04 g) inN,N-dimethylformamide (8 mL) were added potassium phosphate (550 mg) andhydroxylamine hydrochloride (225 mg) at room temperature. The resultingmixture was stirred at 70° C. for 21 h. whereupon the mixture wasdiluted with dichloromethane (20 mL). The insoluble materials areremoved by filtration. The filtrate was concentrated in vacuo and theresidue purified by column chromatography (silica gel, 0-15%methanol/ethyl acetate) to give the title compound (384 mg).

¹H NMR (300 MHz, DMSO-d₆, rotamers present) δ 10.05 (br. s, 1H), 9.77(s, 1H), 7.97-7.94 (m, 2H), 7.78-7.73 (m, 4H), 7.59-7.56 (m, 2H),7.47-7.26 (m, 5H), 5.88 (br. s, 2H), 5.31 (s, 2H), 4.61 (m, 1H),3.67-3.52 (m, 2H), 3.12-3.00 (m, 4H), 2.95-2.72 (m, 5H), 2.62-2.34 (m,4H), 1.76 (m, 1H).

ESI MS m/z 649 (M+H)⁺

Pharmacological Activities

The compounds of the present invention possess factor XIa inhibitoryactivity, for example, such an effect of the compounds of the presentinvention was confirmed by the following tests.

All the procedures were conducted by conventionally used techniques onthe basis of basic biological methods. Furthermore, the measuring methodof the present invention was modified to improve the accuracy and/orsensitivity of measurement for evaluating the compound of the presentinvention. The detailed experimental method was as follows.

Experimental Method

(1) In Vitro Assay

Inhibitory activities of compounds of the present invention againstfactor XIa, Xa, XIIa, IXa, VIIa, plasma kallikrein or thrombin wereevaluated using appropriate purified proteases and synthetic substrates.The rate of hydrolysis of the chromogenic substrate by the relevantprotease was continuously measured at 405 nm. Inhibitory activityagainst each enzyme was calculated as % inhibition using the equationdescribed below.% Inhibition=[[(rate without compound)−(rate with compound)]/(ratewithout compound)]×100%.Each half maximal inhibitory concentration (IC₅₀) value was determinedby plotting the concentration of compound of the invention against the %inhibition.(1-1) Factor XIa Enzyme Activity

Human Factor XIa (Haematologic Technologies Inc.) activity was measuredat an enzyme concentration of 0.1 U/mL in 150 mM NaCl, 5 mM KCl, 1 mg/mLPEG6000, 50 mM HEPES-NaOH (pH7.4) with 300 μM S-2366(pyroGlu-Pro-Arg-pNA, Chromogenix).

(1-2) Plasma Kallikrein Enzyme Activity

Human plasma kallikrein (Enzyme Research Laboratories Ltd) activity wasmeasured at an enzyme concentration of 0.605 mU/mL in 200 mM NaCl, 5mg/mL. PEG6000, 100 mM Phosphate-NaOH (pH7.4) with 150 μM S-2302(H-D-Pro-Phe-Arg-pNA, Chromogenix).

(1-3) Factor Xa and Thrombin Enzyme Activity

Human Factor Xa (American Diagnostica Inc.) and human thrombin (Sigma)activities were measured at the enzyme concentrations of 0.18 U/mL and0.12 U/mL, respectively in the same buffer containing 150 mM NaCl, 2mg/mL PEG6000, 50 mM Tris-HCl (pH7.4), except that the reactions werestarted with 300 μM S-2222 (phenyl-Ile-Glu-Gly-Arg-pNA, Chromogenix) and300 μM S-2366, respectively.

(1-4) Factor XIIa Enzyme Activity

Human Factor α-XIIa (Enzyme Research Laboratories Ltd) activity wasmeasured at an enzyme concentration of 0.17 U/mL in 150 mM NaCl, 50 mMTris-HCl (pH7.4) with 300 μM S-2302 (Pro-Phe-Arg-pNA, Chromogenix).

(1-5) Factor IXa Enzyme Activity

Human Factor IXa (American Diagnostica Inc.) activity was measured at anenzyme concentration of 13 U/mL in 100 mM NaCl, 5 mM CaCl₂, 30% ethyleneglycol, 50 mM Tris-HCl (pH7.4) with 3 mM Pefachrome IXa 3960(Leu-Ph′Gly-Arg-pNA, Pentapharm).

(1-6) Factor VIIa Enzyme Activity

Human Factor VIIa activity was measured using recombinant human factorVIIa (American Diagnostica Inc.) in the presence of recombinant humantissue factor which was produced according to the method described inthe literature (Protein expression and purification, 3, 453-460 (1992)in a buffer containing 150 mM NaCl, 5 mM CaCl₂, 0.5 mg/mL PEG6000, 50 mMHEPES-NaCl (pH7.4) with 3 mM S-2288 (Ile-Pro-Arg-pNA, Chromogenix).

(1-7) APTT, PT Measurement

Activated partial thromboplastin time (APTT) and prothrombin time (PT)were measured using automatic coagulation analyzer (CA-1500, SysmexCorporation). For the APTT or PT measurement, standard human plasma(Siemens Healthcare Diagnostics GmbH) were mixed with each compounddilutions followed by the automatic addition of APTT reagent (SiemensHealthcare Diagnostics GmbH) and 0.02 M calcium chloride or PT reagent(Siemens Healthcare Diagnostics GmbH) to start clot formation. Theanticoagulant activities (APTT2 or PT2) of the compounds of theinvention were expressed as the concentrations necessary to double theclotting time in vehicle (1% DMSO) group. APTT2 or PT2 was determined byplotting the concentration of compound of the invention against the foldincrease of clotting time.

The compounds of the present invention were tested in the factor XIaassay described above, and found to have a good factor XIa inhibitoryactivity as well as good selectivity against other plasma serineproteases. Table I described below lists factor XIa, thrombin and FXaIC₅₀ values measured for the following examples.

TABLE 1 In vitro In vitro In vitro FXIa Thrombin FXa inhibitoryinhibitory inhibitory Example activity activity activity No IC₅₀ (μM)IC₅₀ (μM) IC₅₀ (μM) 9 0.017 >100 >100 20 0.0032 >100 >100 370.013 >100 >100 39 0.078 >100 >100 43 0.011 >100 >100 55 0.014 >100 >10057 0.027 >100 >100 69 0.0074 >100 >100 73 0.0044 37 >100 79 0.006540 >100 86 0.0099 >100 >100 88 0.0054 57 >100 93 0.018 >100 >100 940.0054 >33  >33 96 0.0042 >100 >100 98 0.0023 >100 >100 71 0.002294 >100 99 0.0012 84 >100 111 0.0085 >100 Not tested

Therefore, the results indicated that the compounds of the presentinvention possess factor XIa inhibitory activity as well as highselectivity against other plasma serine proteases.

Additionally, the good oral bioavailability of compounds of the presentinvention can be determined using the following experimental methods.

(2-1) Pharmacokinetic (P) Study in Rat

Each compound of the present invention in a solution of 20% well solve(celeste) was given to fasted male Crj:CD (SD) rats as a single 3 mg/kg,p.o. dose by gavage. Blood samples were drawn from jugular vein intosyringes containing 3.2% sodium citrate (the volume ratio of blood toanticoagulant=9:1) or heparinized syringes at 0.5, 1, 3, 7 hours afteroral administration. Plasma was obtained by centrifugation and stored at−20° C. until measurement of plasma concentration.

To measure plasma concentrations of the compounds of the presentinvention, plasma samples were deproteinized with acetonitrile, followedby evaporation of the acetonitrile to dryness. Then the sample wasreconstituted in the mobile phase and analyzed by LC/MS/MS. Ananalytical column (Shim-pack XR-ODSII, 2.0 mm×75 mm, 2.2 μm) and mobilephase (0.1% formic acid in water and 0.1% formic acid in acetonitrile,flow rate of 0.5 mL/min) were used. The system was used in multiplereaction monitoring (MRM) mode with positive ion detection.

(2-2) Pharmacokinetic (PK) Study of the Compound which has a FunctionalGroup (e.g. an Ester Group, a Substituted Amidine Group, a SubstitutedGuanidine Group, Etc.) in Rat

Each compound of the present invention in a solution of 20% well solve(celeste) was given to fasted male Crj:CD (SD) rats as a single 3 mg/kg,p.o. dose by gavage. Blood samples were drawn from jugular vein intosyringes treated with heparin-diisopropyl fluorophosphate mixture(500:1) at 0.5, 1, 3, 7 hours after oral administration. Plasma wasobtained by centrifugation and stored at −20° C. until measurement ofplasma concentration.

To measure plasma concentrations of the compounds of the presentinvention, plasma samples were deproteinized with acetonitrile, followedby evaporation of the acetonitrile to dryness. Then the sample wasreconstituted in the mobile phase and analyzed by LC/MS/MS. Ananalytical column (Shim-pack XR-ODSII, 2.0 mm×75 mm, 2.2 μm) and mobilephase (0.1% formic acid in water and 0.1% formic acid in acetonitrile,flow rate of 0.5 mL/min) were used. The system was used in multiplereaction monitoring (MRM) mode with positive ion detection.

Additionally, enzymatic hydrolysis of a functional group (e.g. an estergroup, a substituted amidine group, a substituted guanidine group, etc.)in the compound of the present invention can be determined using thefollowing experimental methods.

(3-1) Analysis of Enzymatic Hydrolysis of a Functional Group (e.g. anEster Group, a Substituted Amidine Group, a Substituted Guanidine Group,Etc.) in the Compounds of the Present Invention Using HepatocytesPrepared from Various Species (Rat, Dog, Monkey, Human)

A typical assay procedure was conducted by using cryopreservedhepatocytes prepared from various species. A mixture of hepatocytes,buffer (pH 7.4), and each test compound were incubated. The final testcompound concentration was typically 100 ng/mL, with a usual celldensity of 1,000,000 cells/ml for all species. The incubation was at 37°C., with time-points taken over 120 minutes. Reaction termination wasachieved by addition of an aliquot of the hepatocyte/test compoundmixture to acetonitrile/ethanol (7/3) to effect protein precipitation,followed by centrifugation. Then the sample was diluted with distilledwater and analyzed by LC/MS/MS. An analytical column (Shim-packXR-ODSII, 2.0 mm×75 mm, 2.2 μm) and mobile phase (0.1% formic acid inwater and 0.1% formic acid in acetonitrile, flow rate of 0.5 mL/min)were used. The system was used in multiple reactions monitoring (MRM)mode with positive ion detection.

(3-2) Analysis of Enzymatic Hydrolysis of a Functional Group (e.g. anEster Group, a Substituted Amidine Group, a Substituted Guanidine Group,Etc.) in the Compounds of the Present Invention Using Blood from VariousSpecies (Rat, Dog, Monkey, Human)

Each compound of the present invention in a solution of acetonitrilewere incubated in blood from various species. The incubation wastypically performed at a concentration of 100 ng/mL of test compound at37° C., with time points taken over 60 minutes. The reaction wasterminated by addition of an aliquot of blood/test compound mixture toacetonitrile/ethanol (7/3) to effect protein precipitation, followed bycentrifugation. Then the sample was diluted with distilled water andanalyzed by LC/MS/MS. An analytical column (Shim-pack XR-ODSII, 2.0mm×75 mm, 2.2 μm) and mobile phase (0.1% formic acid in water and 0.1%formic acid in acetonitrile, flow rate of 0.5 mL/min) were used. Thesystem was used in multiple reactions monitoring (MRM) mode withpositive ion detection.

Formulation Example 1

The following components were admixed in conventional method and punchedout to obtain 10,000 tablets each containing 10 mg of active ingredient.

Methyl[4-(4-chloro-2-{(2S,4R)-1-{(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl}-1H-imidazol-5-yl)phenyl]carbamate100 g

Carboxymethylcellulose calcium (disintegrating agent) 20 g

Magnesium stearate (lubricating agent) 10 g

Microcrystalline cellulose 870 g

Formulation Example 2

The following components were admixed in conventional method. Thesolution was sterilized in conventional manner, filtered through dustremoval equipment, placed 5 mL portions into ampoules and sterilized byautoclave to obtain 10,000 ampoules each containing 20 mg of the activeingredient.

Methyl[4-(4-chloro-2-{(2S,4R)-1-(2E)-3-[5-chloro-2-(1H-tetrazol-1-yl)phenyl]-2-propenoyl}-4-[1-(methylsulfonyl)-4-piperidinyl]-2-pyrrolidinyl)-1H-imidazol-5-yl)phenyl]carbamate200 g

mannitol 20 g

distilled water 50 L

INDUSTRIAL APPLICABILITY

The compounds of the present invention represented by formula (I) act aspotent and selective inhibitors of factor XIa without side effects suchas bleeding. In particular, the compounds of the present invention actas Factor XIa inhibitors. Thus the compounds of the present inventionare useful in preventing and/or treating thromboembolic diseases, forexample arterial cardiovascular thromboembolic disorders, venouscardiovascular thromboembolic disorders, arterial cerebrovascularthromboembolic disorders, venous cerebrovascular thromboembolicdisorders and thromboembolic disorders in the chambers of the heart orin the peripheral circulation. The compound of the present invention istherefore useful as a medicament.

The invention claimed is: 1.4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid or a pharmaceutically acceptable salt thereof.
 2. A pharmaceuticalcomposition which comprises4-[({(2S,4S)-1-(4-carbamimidoylbenzoyl)-4-[4-(methylsulfonyl)-1-piperazinyl]-2-pyrrolidinyl}carbonyl)amino]benzoicacid or a pharmaceutically acceptable salt thereof.